CORD-19-Sentences  

CORD-19:005c43980edf3fcc2a4d12ee7ad630ddb651ce6e JSONTXT 8 Projects

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Id Subject Object Predicate Lexical cue
TextSentencer_T1 0-174 Sentence denotes Development of a smartphone-based rapid dual fluorescent diagnostic system for the simultaneous detection of influenza A and H5 subtype in avian influenza A-infected patients
TextSentencer_T1 0-174 Sentence denotes Development of a smartphone-based rapid dual fluorescent diagnostic system for the simultaneous detection of influenza A and H5 subtype in avian influenza A-infected patients
TextSentencer_T2 176-184 Sentence denotes Abstract
TextSentencer_T2 176-184 Sentence denotes Abstract
TextSentencer_T3 185-334 Sentence denotes Accurate and rapid diagnosis of highly pathogenic avian influenza A H5N1 is of critical importance for the effective clinical management of patients.
TextSentencer_T3 185-334 Sentence denotes Accurate and rapid diagnosis of highly pathogenic avian influenza A H5N1 is of critical importance for the effective clinical management of patients.
TextSentencer_T4 335-577 Sentence denotes Here, we developed a rapid and simultaneous detection toolkit for influenza A H5 subtype viruses in human samples based on a bioconjugate of quantum dots (QDs) assembly and a smartphone-based rapid dual fluorescent diagnostic system (SRDFDS).
TextSentencer_T4 335-577 Sentence denotes Here, we developed a rapid and simultaneous detection toolkit for influenza A H5 subtype viruses in human samples based on a bioconjugate of quantum dots (QDs) assembly and a smartphone-based rapid dual fluorescent diagnostic system (SRDFDS).
TextSentencer_T5 578-586 Sentence denotes Methods:
TextSentencer_T5 578-586 Sentence denotes Methods:
TextSentencer_T6 587-744 Sentence denotes Two types of QDs were assembled on a latex bead to enhance the detection sensitivity and specificity of influenza A infection (QD580) and H5 subtype (QD650).
TextSentencer_T6 587-744 Sentence denotes Two types of QDs were assembled on a latex bead to enhance the detection sensitivity and specificity of influenza A infection (QD580) and H5 subtype (QD650).
TextSentencer_T7 745-918 Sentence denotes The dual signals of influenza A and H5 subtype of H5N1-infected patients were detected simultaneously and quantified separately by SRDFDS equipped with two emission filters.
TextSentencer_T7 745-918 Sentence denotes The dual signals of influenza A and H5 subtype of H5N1-infected patients were detected simultaneously and quantified separately by SRDFDS equipped with two emission filters.
TextSentencer_T8 919-927 Sentence denotes Results:
TextSentencer_T8 919-927 Sentence denotes Results:
TextSentencer_T9 928-1124 Sentence denotes Our results showed a high sensitivity of 92.86% (13/14) and 78.57% (11/14), and a specificity of 100% (38/38, P < 0.0001) and 97.37% (37/38) for influenza A and H5 subtype detection, respectively.
TextSentencer_T9 928-1124 Sentence denotes Our results showed a high sensitivity of 92.86% (13/14) and 78.57% (11/14), and a specificity of 100% (38/38, P < 0.0001) and 97.37% (37/38) for influenza A and H5 subtype detection, respectively.
TextSentencer_T10 1125-1136 Sentence denotes Conclusion:
TextSentencer_T10 1125-1136 Sentence denotes Conclusion:
TextSentencer_T11 1137-1367 Sentence denotes Therefore, our multiplex QD bioconjugates and SRDFDS-based influenza virus detection toolkit potentially provide accurate and meaningful diagnosis information with improved detection accuracies and sensitivities for H5N1 patients.
TextSentencer_T11 1137-1367 Sentence denotes Therefore, our multiplex QD bioconjugates and SRDFDS-based influenza virus detection toolkit potentially provide accurate and meaningful diagnosis information with improved detection accuracies and sensitivities for H5N1 patients.
TextSentencer_T12 1369-1497 Sentence denotes Avian influenza (AI) viruses are distinct from human seasonal influenza viruses and are not easily transmissible to humans [1] .
TextSentencer_T12 1369-1497 Sentence denotes Avian influenza (AI) viruses are distinct from human seasonal influenza viruses and are not easily transmissible to humans [1] .
TextSentencer_T13 1498-1652 Sentence denotes However, AI viruses may occasionally infect humans through direct or indirect contact, resulting in disease symptoms ranging from a mild illness to death.
TextSentencer_T13 1498-1652 Sentence denotes However, AI viruses may occasionally infect humans through direct or indirect contact, resulting in disease symptoms ranging from a mild illness to death.
TextSentencer_T14 1653-1732 Sentence denotes AI has become a global concern with a mortality rate of approximately 60% [2] .
TextSentencer_T14 1653-1732 Sentence denotes AI has become a global concern with a mortality rate of approximately 60% [2] .
TextSentencer_T15 1733-1893 Sentence denotes In this context, the H5N1 virus is highly Ivyspring International Publisher contagious among birds and can be deadly, especially in domestic poultry and humans.
TextSentencer_T15 1733-1893 Sentence denotes In this context, the H5N1 virus is highly Ivyspring International Publisher contagious among birds and can be deadly, especially in domestic poultry and humans.
TextSentencer_T16 1894-2123 Sentence denotes Therefore, an on-site surveillance/alert system for the highly pathogenic avian influenza (HPAI) A H5N1 would prevent widespread exposure to the virus and identify potential patients/animals for immediate attention and treatment.
TextSentencer_T16 1894-2123 Sentence denotes Therefore, an on-site surveillance/alert system for the highly pathogenic avian influenza (HPAI) A H5N1 would prevent widespread exposure to the virus and identify potential patients/animals for immediate attention and treatment.
TextSentencer_T17 2124-2240 Sentence denotes Improved case management of diseases with common flu symptoms is especially crucial in densely populated areas [3] .
TextSentencer_T17 2124-2240 Sentence denotes Improved case management of diseases with common flu symptoms is especially crucial in densely populated areas [3] .
TextSentencer_T18 2241-2514 Sentence denotes An adequate H5 subtype-specific diagnostic toolkit for on-site monitoring has not been developed, and current field-level point-of-care testing (POCT) is not recommended for screening AI infections in clinical samples due to insufficient sensitivity and poor accuracy [4] .
TextSentencer_T18 2241-2514 Sentence denotes An adequate H5 subtype-specific diagnostic toolkit for on-site monitoring has not been developed, and current field-level point-of-care testing (POCT) is not recommended for screening AI infections in clinical samples due to insufficient sensitivity and poor accuracy [4] .
TextSentencer_T19 2515-2656 Sentence denotes Simultaneous detection of multiple biomarkers using antigen-specific antibodies can increase the diagnostic accuracy and sensitivity of POCT.
TextSentencer_T19 2515-2656 Sentence denotes Simultaneous detection of multiple biomarkers using antigen-specific antibodies can increase the diagnostic accuracy and sensitivity of POCT.
TextSentencer_T20 2657-2826 Sentence denotes Thus, many researchers have attempted to design a molecular diagnostic method for the simultaneous detection of influenza A and B as well as their subtypes [5] [6] [7] .
TextSentencer_T20 2657-2826 Sentence denotes Thus, many researchers have attempted to design a molecular diagnostic method for the simultaneous detection of influenza A and B as well as their subtypes [5] [6] [7] .
TextSentencer_T21 2827-3080 Sentence denotes Multiplex detection has recently received great attention, and microfluidic devices have emerged as a promising diagnostic platform because of their favorable material transport process, rapid turnaround time, and low sample/reagent consumption [8, 9] .
TextSentencer_T21 2827-3080 Sentence denotes Multiplex detection has recently received great attention, and microfluidic devices have emerged as a promising diagnostic platform because of their favorable material transport process, rapid turnaround time, and low sample/reagent consumption [8, 9] .
TextSentencer_T22 3081-3216 Sentence denotes However, several challenges in mass production, particularly sensor fabrication costs, limit their clinical application for POCT [10] .
TextSentencer_T22 3081-3216 Sentence denotes However, several challenges in mass production, particularly sensor fabrication costs, limit their clinical application for POCT [10] .
TextSentencer_T23 3217-3459 Sentence denotes Given the demand for multiplex testing and the major advantages of the lateral flow assay over alternative rapid diagnostic platforms, this study aimed to adapt the simplicity of the membrane-based lateral flow assay and multiplex techniques.
TextSentencer_T23 3217-3459 Sentence denotes Given the demand for multiplex testing and the major advantages of the lateral flow assay over alternative rapid diagnostic platforms, this study aimed to adapt the simplicity of the membrane-based lateral flow assay and multiplex techniques.
TextSentencer_T24 3460-3643 Sentence denotes Various fluorescent materials have been applied for lateral flow assays as alternative to conventional gold nanoparticles to detect influenza A as POCT [11] [12] [13] [14] [15] [16] .
TextSentencer_T24 3460-3643 Sentence denotes Various fluorescent materials have been applied for lateral flow assays as alternative to conventional gold nanoparticles to detect influenza A as POCT [11] [12] [13] [14] [15] [16] .
TextSentencer_T25 3644-3740 Sentence denotes Also, self-contained and disposable diagnostic devices have been developed [15, [17] [18] [19] .
TextSentencer_T25 3644-3740 Sentence denotes Also, self-contained and disposable diagnostic devices have been developed [15, [17] [18] [19] .
TextSentencer_T26 3741-3873 Sentence denotes There is a critical need for POCT diagnostics in healthcare which can have a positive impact on the quality of life worldwide [20] .
TextSentencer_T26 3741-3873 Sentence denotes There is a critical need for POCT diagnostics in healthcare which can have a positive impact on the quality of life worldwide [20] .
TextSentencer_T27 3874-4049 Sentence denotes POCT devices of interest include smartphones [20] , wearable devices [21] , other complementary metal-oxidesemiconductor (CMOS) imaging devices [22] , and strip readers [23] .
TextSentencer_T27 3874-4049 Sentence denotes POCT devices of interest include smartphones [20] , wearable devices [21] , other complementary metal-oxidesemiconductor (CMOS) imaging devices [22] , and strip readers [23] .
TextSentencer_T28 4050-4432 Sentence denotes Among them, devices integrating with personal computer have revolutionized research in recent decades, and smartphones are now finding increasing application in a wide variety of research domains as they are expedited by the integrated data capture, processing capability (e.g., lab in a phone), operability, compact form, relatively low cost, and very widespread circulation [24] .
TextSentencer_T28 4050-4432 Sentence denotes Among them, devices integrating with personal computer have revolutionized research in recent decades, and smartphones are now finding increasing application in a wide variety of research domains as they are expedited by the integrated data capture, processing capability (e.g., lab in a phone), operability, compact form, relatively low cost, and very widespread circulation [24] .
TextSentencer_T29 4433-4683 Sentence denotes Given that assays combining nanoparticles with consumer electronic devices have almost exclusively utilized optical readout, we assessed the potential for quantum dots (QDs) nanoparticle-based electrochemical assays with readout through a smartphone.
TextSentencer_T29 4433-4683 Sentence denotes Given that assays combining nanoparticles with consumer electronic devices have almost exclusively utilized optical readout, we assessed the potential for quantum dots (QDs) nanoparticle-based electrochemical assays with readout through a smartphone.
TextSentencer_T30 4684-4855 Sentence denotes In particular, QDs have attracted considerable attention because of their improved quantum yield (QY), optical stability, and potential for multiplexing in POCT [25, 26] .
TextSentencer_T30 4684-4855 Sentence denotes In particular, QDs have attracted considerable attention because of their improved quantum yield (QY), optical stability, and potential for multiplexing in POCT [25, 26] .
TextSentencer_T31 4856-4950 Sentence denotes The application of QDs in lateral flow strips via an aptamer is widely used in POCT [27, 28] .
TextSentencer_T31 4856-4950 Sentence denotes The application of QDs in lateral flow strips via an aptamer is widely used in POCT [27, 28] .
TextSentencer_T32 4951-5117 Sentence denotes Despite the versatile tunability of the emission peaks of QDs in multiplexed diagnostic systems, few reports show their application in clinical diagnostics [29, 30] .
TextSentencer_T32 4951-5117 Sentence denotes Despite the versatile tunability of the emission peaks of QDs in multiplexed diagnostic systems, few reports show their application in clinical diagnostics [29, 30] .
TextSentencer_T33 5118-5360 Sentence denotes Since high-fluorescent QDs are usually synthesized at elevated temperatures in organic solvents, they are only soluble in non-polar organic solvents, which are not suitable for biological applications, water compatibility, and bioconjugation.
TextSentencer_T33 5118-5360 Sentence denotes Since high-fluorescent QDs are usually synthesized at elevated temperatures in organic solvents, they are only soluble in non-polar organic solvents, which are not suitable for biological applications, water compatibility, and bioconjugation.
TextSentencer_T34 5361-5566 Sentence denotes When QDs are covered with hydrophilic shells using tedious processing [31] , the signal is too weak for the visible readouts of POCT requiring encapsulation of many QDs in an individual nanoparticle [32] .
TextSentencer_T34 5361-5566 Sentence denotes When QDs are covered with hydrophilic shells using tedious processing [31] , the signal is too weak for the visible readouts of POCT requiring encapsulation of many QDs in an individual nanoparticle [32] .
TextSentencer_T35 5567-5800 Sentence denotes Detection of two influenza A virus subtypes (H5 and H9) via single QD-based immunoassay was previously reported by Wu et al. [33] ; however, it was not simultaneous detection of two antigens of one subtype virus by two different QDs.
TextSentencer_T35 5567-5800 Sentence denotes Detection of two influenza A virus subtypes (H5 and H9) via single QD-based immunoassay was previously reported by Wu et al. [33] ; however, it was not simultaneous detection of two antigens of one subtype virus by two different QDs.
TextSentencer_T36 5801-5949 Sentence denotes Single QD was also successfully used to detect H7N9 by combined magnetic capture, but this was performed under a UV detector rather than POCT [34] .
TextSentencer_T36 5801-5949 Sentence denotes Single QD was also successfully used to detect H7N9 by combined magnetic capture, but this was performed under a UV detector rather than POCT [34] .
TextSentencer_T37 5950-6059 Sentence denotes Also, a laboratory immunoassay for detecting H5N1 using QD was developed and tested in chicken samples [35] .
TextSentencer_T37 5950-6059 Sentence denotes Also, a laboratory immunoassay for detecting H5N1 using QD was developed and tested in chicken samples [35] .
TextSentencer_T38 6060-6212 Sentence denotes Thus, no multiplex POCT diagnostic system for detecting two antigens of a single virus has yet been developed to reliably perform the primary screening.
TextSentencer_T38 6060-6212 Sentence denotes Thus, no multiplex POCT diagnostic system for detecting two antigens of a single virus has yet been developed to reliably perform the primary screening.
TextSentencer_T39 6213-6319 Sentence denotes We had previously developed a portable smartphone-based rapid fluorescent diagnostic system (SRFDS) [13] .
TextSentencer_T39 6213-6319 Sentence denotes We had previously developed a portable smartphone-based rapid fluorescent diagnostic system (SRFDS) [13] .
TextSentencer_T40 6320-6471 Sentence denotes In this study, it was advanced to a multiplex AI virus detection toolkit and readout system equipped with a portable SRDFDS using two emission filters.
TextSentencer_T40 6320-6471 Sentence denotes In this study, it was advanced to a multiplex AI virus detection toolkit and readout system equipped with a portable SRDFDS using two emission filters.
TextSentencer_T41 6472-6670 Sentence denotes The system was designed to simultaneously detect both influenza A and its H5 subtype on a single strip using two QD signals (580 nm and 650 nm) and provides quantitative analysis with high accuracy.
TextSentencer_T41 6472-6670 Sentence denotes The system was designed to simultaneously detect both influenza A and its H5 subtype on a single strip using two QD signals (580 nm and 650 nm) and provides quantitative analysis with high accuracy.
TextSentencer_T42 6671-6792 Sentence denotes This system can be used for the accurate and early diagnosis of influenza viral infections for POCT in clinical settings.
TextSentencer_T42 6671-6792 Sentence denotes This system can be used for the accurate and early diagnosis of influenza viral infections for POCT in clinical settings.
TextSentencer_T43 6793-6862 Sentence denotes Nanobiomaterials require water-soluble QDs of high quality [36, 37] .
TextSentencer_T43 6793-6862 Sentence denotes Nanobiomaterials require water-soluble QDs of high quality [36, 37] .
TextSentencer_T44 6863-7021 Sentence denotes We synthesized water-soluble and stable core-multi-shell QDs (CdSe/CdS/ZnS) using the standard hot injection method with TOPO ligand as the stabilizing agent.
TextSentencer_T44 6863-7021 Sentence denotes We synthesized water-soluble and stable core-multi-shell QDs (CdSe/CdS/ZnS) using the standard hot injection method with TOPO ligand as the stabilizing agent.
TextSentencer_T45 7022-7137 Sentence denotes Subsequently, ligand exchange was carried out to replace the hydrophobic TOPO ligands with hydrophilic MPA ligands.
TextSentencer_T45 7022-7137 Sentence denotes Subsequently, ligand exchange was carried out to replace the hydrophobic TOPO ligands with hydrophilic MPA ligands.
TextSentencer_T46 7138-7311 Sentence denotes The emission wavelength could be tuned by controlling the QD core size, and two types of QDs (QD580 and QD650 at 580 nm and 650 nm emissions, respectively) were synthesized.
TextSentencer_T46 7138-7311 Sentence denotes The emission wavelength could be tuned by controlling the QD core size, and two types of QDs (QD580 and QD650 at 580 nm and 650 nm emissions, respectively) were synthesized.
TextSentencer_T47 7312-7407 Sentence denotes The QYs of the QDs were calculated as 70% and 48% for QD580 and QD650, respectively (Fig. S1) .
TextSentencer_T47 7312-7407 Sentence denotes The QYs of the QDs were calculated as 70% and 48% for QD580 and QD650, respectively (Fig. S1) .
TextSentencer_T48 7408-7501 Sentence denotes The diameters of QD580 and QD650 were 5.4 ± 1.5 nm and 9.3 ± 2.0 nm, respectively (Fig. S2) .
TextSentencer_T48 7408-7501 Sentence denotes The diameters of QD580 and QD650 were 5.4 ± 1.5 nm and 9.3 ± 2.0 nm, respectively (Fig. S2) .
TextSentencer_T49 7502-7696 Sentence denotes This high QY (>70%) has been reported to be measurable under UV light [38] ; however, few studies have investigated the required QYs for a relatively low sensitivity detection tool such as POCT.
TextSentencer_T49 7502-7696 Sentence denotes This high QY (>70%) has been reported to be measurable under UV light [38] ; however, few studies have investigated the required QYs for a relatively low sensitivity detection tool such as POCT.
TextSentencer_T50 7697-8052 Sentence denotes When we observed the fluorescence signals of the QDs under a UV lamp after a strip assay with a high concentration of viruses (160 hemagglutinin units [HAU]/ml) (Fig. 1A) , intensities of both QD580 and QD650 conjugates were inconclusive in both the test and control lines, indicating that QDs alone are not suitable for sensitive virus detection in POCT.
TextSentencer_T50 7697-8052 Sentence denotes When we observed the fluorescence signals of the QDs under a UV lamp after a strip assay with a high concentration of viruses (160 hemagglutinin units [HAU]/ml) (Fig. 1A) , intensities of both QD580 and QD650 conjugates were inconclusive in both the test and control lines, indicating that QDs alone are not suitable for sensitive virus detection in POCT.
TextSentencer_T51 8053-8201 Sentence denotes To amplify the fluorescence intensity, QDs were assembled onto an amine-functionalized latex template using the EDC/NHS coupling method (Fig. 1B) .
TextSentencer_T51 8053-8201 Sentence denotes To amplify the fluorescence intensity, QDs were assembled onto an amine-functionalized latex template using the EDC/NHS coupling method (Fig. 1B) .
TextSentencer_T52 8202-8342 Sentence denotes All remaining amine groups on the latex bead surface were converted to carboxylate groups for conjugation with monoclonal antibodies (mAbs).
TextSentencer_T52 8202-8342 Sentence denotes All remaining amine groups on the latex bead surface were converted to carboxylate groups for conjugation with monoclonal antibodies (mAbs).
TextSentencer_T53 8343-8517 Sentence denotes As shown in Fig. 1C , a large number of hydrophilic MPA-capped CdSe/CdS/ZnS QDs were attached to latex beads, as confirmed by transmission electron microscopy (TEM) analysis.
TextSentencer_T53 8343-8517 Sentence denotes As shown in Fig. 1C , a large number of hydrophilic MPA-capped CdSe/CdS/ZnS QDs were attached to latex beads, as confirmed by transmission electron microscopy (TEM) analysis.
TextSentencer_T54 8518-8768 Sentence denotes Following conjugation of the QDs with mAbs, the sizes of the complexes increased to 185.83 ± 5.25 nm for latex + QD580 + mAb, and 188.38 ± 5.28 nm for latex + QD650 + mAb, compared to the latex alone (175.09 ± 5.27 nm) (n = 30 per sample) (Fig. 1D) .
TextSentencer_T54 8518-8768 Sentence denotes Following conjugation of the QDs with mAbs, the sizes of the complexes increased to 185.83 ± 5.25 nm for latex + QD580 + mAb, and 188.38 ± 5.28 nm for latex + QD650 + mAb, compared to the latex alone (175.09 ± 5.27 nm) (n = 30 per sample) (Fig. 1D) .
TextSentencer_T55 8769-8917 Sentence denotes This diameter of the latex was chosen because it was relatively bigger than the antibody (hydrodynamic diameter of ∼10 nm) [39] and the QDs (10 nm).
TextSentencer_T55 8769-8917 Sentence denotes This diameter of the latex was chosen because it was relatively bigger than the antibody (hydrodynamic diameter of ∼10 nm) [39] and the QDs (10 nm).
TextSentencer_T56 8918-9125 Sentence denotes Empirically, a latex larger than 175 nm caused precipitation on the membrane thereby hindering the lateral flow, and smaller than 100 nm latex could not efficiently carry the antibody and QDs on the surface.
TextSentencer_T56 8918-9125 Sentence denotes Empirically, a latex larger than 175 nm caused precipitation on the membrane thereby hindering the lateral flow, and smaller than 100 nm latex could not efficiently carry the antibody and QDs on the surface.
TextSentencer_T57 9126-9449 Sentence denotes According to our computational calculations of the average diameter of the QDs, latex, and length of MPA ligands, the surface of the latex was conjugated with an average of 2,243 numbers of QD580 and 924 of QD650 beads per latex, respectively (details of this calculation are shown in Supplementary Information Scheme S1) .
TextSentencer_T57 9126-9449 Sentence denotes According to our computational calculations of the average diameter of the QDs, latex, and length of MPA ligands, the surface of the latex was conjugated with an average of 2,243 numbers of QD580 and 924 of QD650 beads per latex, respectively (details of this calculation are shown in Supplementary Information Scheme S1) .
TextSentencer_T58 9450-9592 Sentence denotes Each latex bioconjugate was tested using a rapid fluorescent immunochromatographic assay, and fluorescent images were captured under UV light.
TextSentencer_T58 9450-9592 Sentence denotes Each latex bioconjugate was tested using a rapid fluorescent immunochromatographic assay, and fluorescent images were captured under UV light.
TextSentencer_T59 9593-9696 Sentence denotes Pixels were counted using ImageJ software to determine the dynamic range of quantification (Fig. 1E) .
TextSentencer_T59 9593-9696 Sentence denotes Pixels were counted using ImageJ software to determine the dynamic range of quantification (Fig. 1E) .
TextSentencer_T60 9697-9988 Sentence denotes The ImageJ software was used to interpret the raw data in two dimensions with the length and width of the band defined by the repeated 'rectangle tool' such that the chemiluminescent signal emitted from the band was registered as a peak rising out of the test line (TL) or control line (CL).
TextSentencer_T60 9697-9988 Sentence denotes The ImageJ software was used to interpret the raw data in two dimensions with the length and width of the band defined by the repeated 'rectangle tool' such that the chemiluminescent signal emitted from the band was registered as a peak rising out of the test line (TL) or control line (CL).
TextSentencer_T61 9989-10128 Sentence denotes In this study, each TL was subtracted from the background and was normalized by the ratio of the density of CL to calculate TL/CL in Fig 1.
TextSentencer_T61 9989-10128 Sentence denotes In this study, each TL was subtracted from the background and was normalized by the ratio of the density of CL to calculate TL/CL in Fig 1.
TextSentencer_T62 10129-10336 Sentence denotes As shown in Fig. 1F , both latex bioconjugates had a high linear regression (QD580 conjugate: r 2 = 0.9706, and QD650 conjugate: r 2 = 0.9649) for the H5N3 virus at concentrations of 5-160 HAU/mL (Fig. S3) .
TextSentencer_T62 10129-10336 Sentence denotes As shown in Fig. 1F , both latex bioconjugates had a high linear regression (QD580 conjugate: r 2 = 0.9706, and QD650 conjugate: r 2 = 0.9649) for the H5N3 virus at concentrations of 5-160 HAU/mL (Fig. S3) .
TextSentencer_T63 10337-10539 Sentence denotes To determine the presence of AI virus antigens on the strips, image acquisition and fluorescence intensity estimation were performed on two TLs and a CL on a lateral flow immunoassay strip using SRDFDS.
TextSentencer_T63 10337-10539 Sentence denotes To determine the presence of AI virus antigens on the strips, image acquisition and fluorescence intensity estimation were performed on two TLs and a CL on a lateral flow immunoassay strip using SRDFDS.
TextSentencer_T64 10540-10739 Sentence denotes Two bioconjugates were simultaneously used with SRDFDS to achieve high fluorescence detection sensitivity within a single strip and to analyze a separate lateral flow reaction accurately ( Fig. 2A) .
TextSentencer_T64 10540-10739 Sentence denotes Two bioconjugates were simultaneously used with SRDFDS to achieve high fluorescence detection sensitivity within a single strip and to analyze a separate lateral flow reaction accurately ( Fig. 2A) .
TextSentencer_T65 10740-10966 Sentence denotes The fluorescence intensity of the two QDs bioconjugates was measured by a smartphone detector utilizing the same platform containing a pair of parabolic mirrors between the lateral flow strip and the smartphone's camera [13] .
TextSentencer_T65 10740-10966 Sentence denotes The fluorescence intensity of the two QDs bioconjugates was measured by a smartphone detector utilizing the same platform containing a pair of parabolic mirrors between the lateral flow strip and the smartphone's camera [13] .
TextSentencer_T66 10967-11168 Sentence denotes However, current SRDFDS was designed to get fluorescent signals of two QDs separately in the CL, using two different emission filters (580 nm and 650 nm) which were inserted into the module (Fig. 2B) .
TextSentencer_T66 10967-11168 Sentence denotes However, current SRDFDS was designed to get fluorescent signals of two QDs separately in the CL, using two different emission filters (580 nm and 650 nm) which were inserted into the module (Fig. 2B) .
TextSentencer_T67 11169-11303 Sentence denotes In the lateral flow, a control line was used to confirm completion of the testing reaction by capturing the antibody on the conjugate.
TextSentencer_T67 11169-11303 Sentence denotes In the lateral flow, a control line was used to confirm completion of the testing reaction by capturing the antibody on the conjugate.
TextSentencer_T68 11304-11377 Sentence denotes Our study was performed using both conjugate mouse monoclonal antibodies.
TextSentencer_T68 11304-11377 Sentence denotes Our study was performed using both conjugate mouse monoclonal antibodies.
TextSentencer_T69 11378-11530 Sentence denotes As the species of anti-influenza A NP and H5 subtype were the same, anti-mouse IgGs coated on one control line were expected to capture both conjugates.
TextSentencer_T69 11378-11530 Sentence denotes As the species of anti-influenza A NP and H5 subtype were the same, anti-mouse IgGs coated on one control line were expected to capture both conjugates.
TextSentencer_T70 11531-11654 Sentence denotes With emission filters, the fluorescent signal of each conjugate captured on the control line could be obtained, separately.
TextSentencer_T70 11531-11654 Sentence denotes With emission filters, the fluorescent signal of each conjugate captured on the control line could be obtained, separately.
TextSentencer_T71 11655-11916 Sentence denotes The emission filter at 580 nm had 585 nm wavelength (at center) and 65 nm bandwidth, while the emission filter at 650 nm had 650 nm wavelength (at center) and 45 nm bandwidth, indicating that the two emission filters could discriminate each QD signal at one CL.
TextSentencer_T71 11655-11916 Sentence denotes The emission filter at 580 nm had 585 nm wavelength (at center) and 65 nm bandwidth, while the emission filter at 650 nm had 650 nm wavelength (at center) and 45 nm bandwidth, indicating that the two emission filters could discriminate each QD signal at one CL.
TextSentencer_T72 11917-12003 Sentence denotes The smartphone-based fluorescence detector is presented on the right side of Fig. 2B .
TextSentencer_T72 11917-12003 Sentence denotes The smartphone-based fluorescence detector is presented on the right side of Fig. 2B .
TextSentencer_T73 12004-12192 Sentence denotes To measure each fluorescence value under different emission filters, the fluorescence intensity values of each line were measured after changing the position of the line with a micrometer.
TextSentencer_T73 12004-12192 Sentence denotes To measure each fluorescence value under different emission filters, the fluorescence intensity values of each line were measured after changing the position of the line with a micrometer.
TextSentencer_T74 12193-12275 Sentence denotes The LED current controller regulated the energy level independent of the detector.
TextSentencer_T74 12193-12275 Sentence denotes The LED current controller regulated the energy level independent of the detector.
TextSentencer_T75 12276-12596 Sentence denotes As described previously, an application running on the Android operating system was developed to enable non-experts to easily complete a fluorescence immunochromatographic diagnostic test and report its results via wireless communication through step-by-step instructions displayed on the smartphone's touchscreen [13] .
TextSentencer_T75 12276-12596 Sentence denotes As described previously, an application running on the Android operating system was developed to enable non-experts to easily complete a fluorescence immunochromatographic diagnostic test and report its results via wireless communication through step-by-step instructions displayed on the smartphone's touchscreen [13] .
TextSentencer_T76 12597-12729 Sentence denotes After completing the measurements, the test result was obtained by computing the ratio of fluorescence intensity (TL1/CL or TL2/CL).
TextSentencer_T76 12597-12729 Sentence denotes After completing the measurements, the test result was obtained by computing the ratio of fluorescence intensity (TL1/CL or TL2/CL).
TextSentencer_T77 12730-12936 Sentence denotes To achieve a simultaneous diagnostic performance, dual quantum dots-mediated separate fluorescence signals for each test line (TL) and control line (CL) with two different emission filters must be measured.
TextSentencer_T77 12730-12936 Sentence denotes To achieve a simultaneous diagnostic performance, dual quantum dots-mediated separate fluorescence signals for each test line (TL) and control line (CL) with two different emission filters must be measured.
TextSentencer_T78 12937-13192 Sentence denotes SRDFDS can normalize the positive sample compared to the negative sample using each value of TL/CL [TL1/CL for H5-HA detection with 580 nm emission filter (F580 nm) and TL2/CL for influenza A with 650 nm emission filter (F650 nm)] and the fold of control.
TextSentencer_T78 12937-13192 Sentence denotes SRDFDS can normalize the positive sample compared to the negative sample using each value of TL/CL [TL1/CL for H5-HA detection with 580 nm emission filter (F580 nm) and TL2/CL for influenza A with 650 nm emission filter (F650 nm)] and the fold of control.
TextSentencer_T79 13193-13346 Sentence denotes Finally, all results obtained by SRDFDS can be analyzed with dual fluorescence signals leading to accurate diagnostic result using only TL1/CL or TL2/CL.
TextSentencer_T79 13193-13346 Sentence denotes Finally, all results obtained by SRDFDS can be analyzed with dual fluorescence signals leading to accurate diagnostic result using only TL1/CL or TL2/CL.
TextSentencer_T80 13347-13412 Sentence denotes Fig. 3A shows the strategy of signal analyses measured by SRDFDS.
TextSentencer_T80 13347-13412 Sentence denotes Fig. 3A shows the strategy of signal analyses measured by SRDFDS.
TextSentencer_T81 13413-13657 Sentence denotes The TL2/CL value at 580 nm and TL1/CL value at 650 nm of the positive sample (ii in Figure 3A ) were divided by the corresponding values of the uninfected sample (i in Figure 3A ) to normalize and obtain the final TL/CL value (fold of control).
TextSentencer_T81 13413-13657 Sentence denotes The TL2/CL value at 580 nm and TL1/CL value at 650 nm of the positive sample (ii in Figure 3A ) were divided by the corresponding values of the uninfected sample (i in Figure 3A ) to normalize and obtain the final TL/CL value (fold of control).
TextSentencer_T82 13658-13792 Sentence denotes The TL/CL values of the counterpart test lines (iii and iv) were also obtained to check cross-reactions and further increase accuracy.
TextSentencer_T82 13658-13792 Sentence denotes The TL/CL values of the counterpart test lines (iii and iv) were also obtained to check cross-reactions and further increase accuracy.
TextSentencer_T83 13793-13899 Sentence denotes As shown in Table 1 , inter-assay coefficients of variation (CV) were less than 11 % for all TC/CL values.
TextSentencer_T83 13793-13899 Sentence denotes As shown in Table 1 , inter-assay coefficients of variation (CV) were less than 11 % for all TC/CL values.
TextSentencer_T84 13900-14080 Sentence denotes Following SRDFDS measurements, all diagnostic strips were observed under UV excitation with or without two emission filters to further confirm the accuracy of the assay (Fig. 3B) .
TextSentencer_T84 13900-14080 Sentence denotes Following SRDFDS measurements, all diagnostic strips were observed under UV excitation with or without two emission filters to further confirm the accuracy of the assay (Fig. 3B) .
TextSentencer_T85 14082-14312 Sentence denotes For developing a dual diagnostic system, it was essential to optimize the fluorescent immunochromatographic strip test (FICT) conditions to prevent cross-reactivity of two test lines with two target antigens on one viral particle.
TextSentencer_T85 14082-14312 Sentence denotes For developing a dual diagnostic system, it was essential to optimize the fluorescent immunochromatographic strip test (FICT) conditions to prevent cross-reactivity of two test lines with two target antigens on one viral particle.
TextSentencer_T86 14313-14409 Sentence denotes For accurate normalization of each TL/CL, both conjugates must interact with only one test line.
TextSentencer_T86 14313-14409 Sentence denotes For accurate normalization of each TL/CL, both conjugates must interact with only one test line.
TextSentencer_T87 14410-14472 Sentence denotes CL indicates the leftover conjugate amount after lateral flow.
TextSentencer_T87 14410-14472 Sentence denotes CL indicates the leftover conjugate amount after lateral flow.
TextSentencer_T88 14473-14626 Sentence denotes If some fraction of the conjugate is randomly captured by the counter test line, its concentration would be decreased thereby increasing the TL/CL value.
TextSentencer_T88 14473-14626 Sentence denotes If some fraction of the conjugate is randomly captured by the counter test line, its concentration would be decreased thereby increasing the TL/CL value.
TextSentencer_T89 14627-14696 Sentence denotes Therefore, in the case of cross-reaction, a high TL/CL is inaccurate.
TextSentencer_T89 14627-14696 Sentence denotes Therefore, in the case of cross-reaction, a high TL/CL is inaccurate.
TextSentencer_T90 14697-14873 Sentence denotes For one virus particle to specifically bind to the bioconjugate in each TL, all viral antigens need to be released making the composition of the lysis buffer a critical factor.
TextSentencer_T90 14697-14873 Sentence denotes For one virus particle to specifically bind to the bioconjugate in each TL, all viral antigens need to be released making the composition of the lysis buffer a critical factor.
TextSentencer_T91 14874-15069 Sentence denotes We, therefore, tested different concentrations of sodium dodecyl sulfate (SDS) at different pH values to determine the suitable range for a specific reaction without interference by the other TL.
TextSentencer_T91 14874-15069 Sentence denotes We, therefore, tested different concentrations of sodium dodecyl sulfate (SDS) at different pH values to determine the suitable range for a specific reaction without interference by the other TL.
TextSentencer_T92 15070-15291 Sentence denotes As displayed in Fig. 4 , the lysis buffer was tested with different concentrations of SDS (0.6, 1.2, and 1.5%) at different pH values (5, 8, and 11) using 100-fold dilution of both conjugates with H5N3 virus (640 HAU/mL).
TextSentencer_T92 15070-15291 Sentence denotes As displayed in Fig. 4 , the lysis buffer was tested with different concentrations of SDS (0.6, 1.2, and 1.5%) at different pH values (5, 8, and 11) using 100-fold dilution of both conjugates with H5N3 virus (640 HAU/mL).
TextSentencer_T93 15292-15461 Sentence denotes As the rapid diagnostic strip was not ELISA-based and lacked multiple washing steps, it readily showed nonspecific reactions based on the characteristics of an antibody.
TextSentencer_T93 15292-15461 Sentence denotes As the rapid diagnostic strip was not ELISA-based and lacked multiple washing steps, it readily showed nonspecific reactions based on the characteristics of an antibody.
TextSentencer_T94 15462-15632 Sentence denotes The H5 HA-specific antibody at TL1 cross-reacted with the influenza A antibody conjugate (QD580) in 0.6% SDS at pH 5.0 which was reduced at higher pH values (Fig. 4A-C) .
TextSentencer_T94 15462-15632 Sentence denotes The H5 HA-specific antibody at TL1 cross-reacted with the influenza A antibody conjugate (QD580) in 0.6% SDS at pH 5.0 which was reduced at higher pH values (Fig. 4A-C) .
TextSentencer_T95 15633-15764 Sentence denotes The QD580 conjugate showed a non-specific reaction at TL1 regardless of the presence of the virus affecting TL/CL at normalization.
TextSentencer_T95 15633-15764 Sentence denotes The QD580 conjugate showed a non-specific reaction at TL1 regardless of the presence of the virus affecting TL/CL at normalization.
TextSentencer_T96 15765-15899 Sentence denotes The reduced amount of CL was due to the waste of the conjugate at the other test line rendering the normalization of TL/CL inaccurate.
TextSentencer_T96 15765-15899 Sentence denotes The reduced amount of CL was due to the waste of the conjugate at the other test line rendering the normalization of TL/CL inaccurate.
TextSentencer_T97 15900-16079 Sentence denotes As 0.6% SDS in lysis buffer (pH 11.0) (Fig. 4C) was not sufficient in removing the cross signaling at TL1, we increased the SDS concentration at different pH values ( Fig. 4D-F) .
TextSentencer_T97 15900-16079 Sentence denotes As 0.6% SDS in lysis buffer (pH 11.0) (Fig. 4C) was not sufficient in removing the cross signaling at TL1, we increased the SDS concentration at different pH values ( Fig. 4D-F) .
TextSentencer_T98 16080-16258 Sentence denotes As displayed in Fig. 4F , compared with 1.5% SDS (Fig. 4G-4I) , the lysis buffer with 1.2% SDS at pH 11.0 led to a more consistent TL/CL value at each TL without cross signaling.
TextSentencer_T98 16080-16258 Sentence denotes As displayed in Fig. 4F , compared with 1.5% SDS (Fig. 4G-4I) , the lysis buffer with 1.2% SDS at pH 11.0 led to a more consistent TL/CL value at each TL without cross signaling.
TextSentencer_T99 16259-16455 Sentence denotes Therefore, of the nine different combinations of SDS and pH tested, 1.2% SDS at pH 11.0 was most effective in suppressing the cross-reaction of both QD bioconjugates against counterpart test line.
TextSentencer_T99 16259-16455 Sentence denotes Therefore, of the nine different combinations of SDS and pH tested, 1.2% SDS at pH 11.0 was most effective in suppressing the cross-reaction of both QD bioconjugates against counterpart test line.
TextSentencer_T100 16456-16658 Sentence denotes The performance of the current smartphone-based diagnostic system was, therefore, further optimized to increase the TL/CL ratio by adjusting the two variables in the lysis buffer (1.2% SDS and pH 11.0).
TextSentencer_T100 16456-16658 Sentence denotes The performance of the current smartphone-based diagnostic system was, therefore, further optimized to increase the TL/CL ratio by adjusting the two variables in the lysis buffer (1.2% SDS and pH 11.0).
TextSentencer_T101 16659-16704 Sentence denotes All raw SRDFDS results are shown in Fig. S4 .
TextSentencer_T101 16659-16704 Sentence denotes All raw SRDFDS results are shown in Fig. S4 .
TextSentencer_T102 16705-16966 Sentence denotes To increase the TL/CL ratio, different dilutions (50, 70 , and 100 folds) of QD580-conjugated anti-influenza A NP and QD650-conjugated anti-H5 HA antibody were applied to SRDFDS in the optimized lysis buffer (1.2% SDS and pH 11.0) in the following combinations:
TextSentencer_T102 16705-16966 Sentence denotes To increase the TL/CL ratio, different dilutions (50, 70 , and 100 folds) of QD580-conjugated anti-influenza A NP and QD650-conjugated anti-H5 HA antibody were applied to SRDFDS in the optimized lysis buffer (1.2% SDS and pH 11.0) in the following combinations:
TextSentencer_T103 16967-17170 Sentence denotes The 50-, 70-, and 100-fold diluted QD650 conjugate (to detect H5 HA) was mixed with differentially diluted QD580 conjugate (to detect influenza A NP) with 50-, 70-, and 100-fold dilutions ( Fig. 5A-5I ).
TextSentencer_T103 16967-17170 Sentence denotes The 50-, 70-, and 100-fold diluted QD650 conjugate (to detect H5 HA) was mixed with differentially diluted QD580 conjugate (to detect influenza A NP) with 50-, 70-, and 100-fold dilutions ( Fig. 5A-5I ).
TextSentencer_T104 17171-17522 Sentence denotes The highest concentrations of bioconjugates (H5 HA conjugate: influenza A conjugate=50:50) not only interfered with migration on the strips, but also increased the relative CL value decreasing the TL/CL ratio, indicating that there is no significant difference between the negative sample and H5N3 (20 HAU/mL) in terms of 'fold of control' (Fig. 5A) .
TextSentencer_T104 17171-17522 Sentence denotes The highest concentrations of bioconjugates (H5 HA conjugate: influenza A conjugate=50:50) not only interfered with migration on the strips, but also increased the relative CL value decreasing the TL/CL ratio, indicating that there is no significant difference between the negative sample and H5N3 (20 HAU/mL) in terms of 'fold of control' (Fig. 5A) .
TextSentencer_T105 17523-17792 Sentence denotes As shown in Fig. 5B -5E, at 20 HAU/mL of H5N3 virus, the following combinations of the dilutions of QD650-conjugated anti-H5 HA antibody: QD580-conjugated anti-influenza A NP showed a significant difference without any cross-reactions in the TL/CL value of counterpart:
TextSentencer_T105 17523-17792 Sentence denotes As shown in Fig. 5B -5E, at 20 HAU/mL of H5N3 virus, the following combinations of the dilutions of QD650-conjugated anti-H5 HA antibody: QD580-conjugated anti-influenza A NP showed a significant difference without any cross-reactions in the TL/CL value of counterpart:
TextSentencer_T106 17793-17906 Sentence denotes 50-fold:70-fold (P < 0.001); 50-fold:100-fold (P < 0.05); 70-fold:50-fold (P < 0.05); 70-fold:70-fold (P < 0.01).
TextSentencer_T106 17793-17906 Sentence denotes 50-fold:70-fold (P < 0.001); 50-fold:100-fold (P < 0.05); 70-fold:50-fold (P < 0.05); 70-fold:70-fold (P < 0.01).
TextSentencer_T107 17907-18057 Sentence denotes Further study was performed with diluents of 50-fold and 70-fold of QD650-conjugated anti-H5 HA and QD580conjugated anti-influenza A NP, respectively.
TextSentencer_T107 17907-18057 Sentence denotes Further study was performed with diluents of 50-fold and 70-fold of QD650-conjugated anti-H5 HA and QD580conjugated anti-influenza A NP, respectively.
TextSentencer_T108 18058-18103 Sentence denotes All raw SRDFDS results are shown in Fig. S5 .
TextSentencer_T108 18058-18103 Sentence denotes All raw SRDFDS results are shown in Fig. S5 .
TextSentencer_T109 18104-18324 Sentence denotes After establishing the ideal conditions for the bioconjugate to detect both influenza A and HA on the strip, we examined the quantitative range of SRDFDS for detecting the H5N3 virus using a serial dilution of the virus.
TextSentencer_T109 18104-18324 Sentence denotes After establishing the ideal conditions for the bioconjugate to detect both influenza A and HA on the strip, we examined the quantitative range of SRDFDS for detecting the H5N3 virus using a serial dilution of the virus.
TextSentencer_T110 18325-18458 Sentence denotes Representative images taken using SRDFDS and UV were compared in the presence of the two bioconjugates and are displayed in Fig. 6A .
TextSentencer_T110 18325-18458 Sentence denotes Representative images taken using SRDFDS and UV were compared in the presence of the two bioconjugates and are displayed in Fig. 6A .
TextSentencer_T111 18459-18630 Sentence denotes The fluorescence signals of each TL increased at their respective wavelengths for concentrations of up to 160 HAU/mL proportional to the virus titer by both SRDFDS and UV.
TextSentencer_T111 18459-18630 Sentence denotes The fluorescence signals of each TL increased at their respective wavelengths for concentrations of up to 160 HAU/mL proportional to the virus titer by both SRDFDS and UV.
TextSentencer_T112 18631-18719 Sentence denotes At 160 HAU/mL, TL2 of the influenza A showed cross-reaction at F650 (P < 0.05) under UV.
TextSentencer_T112 18631-18719 Sentence denotes At 160 HAU/mL, TL2 of the influenza A showed cross-reaction at F650 (P < 0.05) under UV.
TextSentencer_T113 18720-18957 Sentence denotes On the contrary, we did not identify any cross-reactivity of QD650-conjugated anti-H5 HA at TL1 of H5 HA at F580, indicating that this emission filter (F580) could discriminate the fluorescence signal of TL1 derived from QD650 conjugate.
TextSentencer_T113 18720-18957 Sentence denotes On the contrary, we did not identify any cross-reactivity of QD650-conjugated anti-H5 HA at TL1 of H5 HA at F580, indicating that this emission filter (F580) could discriminate the fluorescence signal of TL1 derived from QD650 conjugate.
TextSentencer_T114 18958-19066 Sentence denotes We determined the limit of detection (LOD) in our immunoassay based on the statistics shown in Fig. 6B -6E.
TextSentencer_T114 18958-19066 Sentence denotes We determined the limit of detection (LOD) in our immunoassay based on the statistics shown in Fig. 6B -6E.
TextSentencer_T115 19067-19159 Sentence denotes Using F580, the virus titer corresponding to the LOD of influenza A NP was 10 HAU/mL [TL/CL:
TextSentencer_T115 19067-19159 Sentence denotes Using F580, the virus titer corresponding to the LOD of influenza A NP was 10 HAU/mL [TL/CL:
TextSentencer_T116 19160-19196 Sentence denotes 1.80 ± 0.13 (mean ± SD)] (P < 0.05).
TextSentencer_T116 19160-19196 Sentence denotes 1.80 ± 0.13 (mean ± SD)] (P < 0.05).
TextSentencer_T117 19197-19276 Sentence denotes The virus titer corresponding to the LOD of H5 HA at F650 was 10 HAU/mL [TL/CL:
TextSentencer_T117 19197-19276 Sentence denotes The virus titer corresponding to the LOD of H5 HA at F650 was 10 HAU/mL [TL/CL:
TextSentencer_T118 19277-19313 Sentence denotes 1.77 ± 0.24 (mean ± SD)] (P < 0.05).
TextSentencer_T118 19277-19313 Sentence denotes 1.77 ± 0.24 (mean ± SD)] (P < 0.05).
TextSentencer_T119 19314-19520 Sentence denotes The LODs obtained from multiplexed QD conjugates for different emission filters were compared to those with one bioconjugate and a negative control conjugate (one antibody was replaced with anti-mouse IgG).
TextSentencer_T119 19314-19520 Sentence denotes The LODs obtained from multiplexed QD conjugates for different emission filters were compared to those with one bioconjugate and a negative control conjugate (one antibody was replaced with anti-mouse IgG).
TextSentencer_T120 19521-19627 Sentence denotes The LOD of the single conjugate was still 10 HAU/mL for both influenza A (P < 0.05) and H5 HA (P < 0.001).
TextSentencer_T120 19521-19627 Sentence denotes The LOD of the single conjugate was still 10 HAU/mL for both influenza A (P < 0.05) and H5 HA (P < 0.001).
TextSentencer_T121 19628-19888 Sentence denotes At the high titer of 160 HAU/mL, there were no cross signals from the conjugates at both emission filters, indicating that at this high virus titer, there was no random binding of the conjugate due to cross-signaling or loss of conjugate on the counterpart TL.
TextSentencer_T121 19628-19888 Sentence denotes At the high titer of 160 HAU/mL, there were no cross signals from the conjugates at both emission filters, indicating that at this high virus titer, there was no random binding of the conjugate due to cross-signaling or loss of conjugate on the counterpart TL.
TextSentencer_T122 19889-20131 Sentence denotes It implied that the problem of optical leaking at F650 used in SRDFDS could be ignored in computing TL/CL because the lack of random binding of the conjugate to test lines and non-wasted conjugate leftover resulted in an accurate TL/CL value.
TextSentencer_T122 19889-20131 Sentence denotes It implied that the problem of optical leaking at F650 used in SRDFDS could be ignored in computing TL/CL because the lack of random binding of the conjugate to test lines and non-wasted conjugate leftover resulted in an accurate TL/CL value.
TextSentencer_T123 20132-20216 Sentence denotes The negative control conjugates showed a clear reaction from lateral flow in SRDFDS.
TextSentencer_T123 20132-20216 Sentence denotes The negative control conjugates showed a clear reaction from lateral flow in SRDFDS.
TextSentencer_T124 20217-20262 Sentence denotes All raw SRDFDS results are shown in Fig. S6 .
TextSentencer_T124 20217-20262 Sentence denotes All raw SRDFDS results are shown in Fig. S6 .
TextSentencer_T125 20263-20404 Sentence denotes The RNA copy number of the LOD (10 HAU/mL) for SRDFDS was determined by reverse transcription-polymerase chain reaction (rRT-PCR) (Fig. S7 ).
TextSentencer_T125 20263-20404 Sentence denotes The RNA copy number of the LOD (10 HAU/mL) for SRDFDS was determined by reverse transcription-polymerase chain reaction (rRT-PCR) (Fig. S7 ).
TextSentencer_T126 20405-20706 Sentence denotes The LOD of the fluorescent immunochromatographic strip test (FICT; 10 HAU/mL) corresponded to a Ct value of 27.86 ± 0.26 (mean ± SD) and 3.3 × 10 5 ± 5.2 × 10 3 (mean ± SD) RNA/µL for influenza A NP, and a Ct value of 28.27 ± 0.095 (mean ± SD) and 5.9 × 10 5 ± 3.9 × 10 3 (mean ± SD) RNA/µL for H5 HA.
TextSentencer_T126 20405-20706 Sentence denotes The LOD of the fluorescent immunochromatographic strip test (FICT; 10 HAU/mL) corresponded to a Ct value of 27.86 ± 0.26 (mean ± SD) and 3.3 × 10 5 ± 5.2 × 10 3 (mean ± SD) RNA/µL for influenza A NP, and a Ct value of 28.27 ± 0.095 (mean ± SD) and 5.9 × 10 5 ± 3.9 × 10 3 (mean ± SD) RNA/µL for H5 HA.
TextSentencer_T127 20707-20814 Sentence denotes The accuracy of SRDFDS was examined by computing the inter-assay coefficient of variation (CV) ( Table 1) .
TextSentencer_T127 20707-20814 Sentence denotes The accuracy of SRDFDS was examined by computing the inter-assay coefficient of variation (CV) ( Table 1) .
TextSentencer_T128 20815-20970 Sentence denotes The virus titer of spiked specimens was adjusted to 40 or 160 HAU/mL by mixing 10 µL of the appropriate virus stock with 90 µL of nasopharyngeal specimens.
TextSentencer_T128 20815-20970 Sentence denotes The virus titer of spiked specimens was adjusted to 40 or 160 HAU/mL by mixing 10 µL of the appropriate virus stock with 90 µL of nasopharyngeal specimens.
TextSentencer_T129 20971-21053 Sentence denotes The ratios of TL/CL derived from all experiments were then plotted for comparison.
TextSentencer_T129 20971-21053 Sentence denotes The ratios of TL/CL derived from all experiments were then plotted for comparison.
TextSentencer_T130 21054-21247 Sentence denotes TL/CL between the negative control and specimens with the virus at 40 HAU/mL or 160 HAU/mL showed a significant difference for influenza A and H5 HA at the relevant emission filter (P < 0.001).
TextSentencer_T130 21054-21247 Sentence denotes TL/CL between the negative control and specimens with the virus at 40 HAU/mL or 160 HAU/mL showed a significant difference for influenza A and H5 HA at the relevant emission filter (P < 0.001).
TextSentencer_T131 21248-21357 Sentence denotes The analytical error rate was computed by the inter-assay CV based on replicates of 40 HAU/mL and 160 HAU/mL.
TextSentencer_T131 21248-21357 Sentence denotes The analytical error rate was computed by the inter-assay CV based on replicates of 40 HAU/mL and 160 HAU/mL.
TextSentencer_T132 21358-21593 Sentence denotes The inter-assay CV of SRDFDS for influenza A detection using F580 was 6.10%, and that of H5 HA detection using F650 was 9.14%, demonstrating that SRDFDS is an accurate technique because the range of inter-assay CVs was below 15% [40] .
TextSentencer_T132 21358-21593 Sentence denotes The inter-assay CV of SRDFDS for influenza A detection using F580 was 6.10%, and that of H5 HA detection using F650 was 9.14%, demonstrating that SRDFDS is an accurate technique because the range of inter-assay CVs was below 15% [40] .
TextSentencer_T133 21594-21639 Sentence denotes All raw SRDFDS results are shown in Fig. S8 .
TextSentencer_T133 21594-21639 Sentence denotes All raw SRDFDS results are shown in Fig. S8 .
TextSentencer_T134 21640-21750 Sentence denotes Intraday CV was measured by calculating the relative standard deviation (RSD) between 20 and 160 HAU/mL [41] .
TextSentencer_T134 21640-21750 Sentence denotes Intraday CV was measured by calculating the relative standard deviation (RSD) between 20 and 160 HAU/mL [41] .
TextSentencer_T135 21751-21994 Sentence denotes RSD of influenza A was 2.798 ± 0.7 % (mean ± SD) and that of H5 was 4.73 ± 1.3 % (mean ± SD), indicating that this assay was reliable (Fig. S9) . (50, 75 , and 100) were tested in lysis buffer (1.2% SDS and pH 11.0) at 20 HAU/mL of H5N3 virus.
TextSentencer_T135 21751-21994 Sentence denotes RSD of influenza A was 2.798 ± 0.7 % (mean ± SD) and that of H5 was 4.73 ± 1.3 % (mean ± SD), indicating that this assay was reliable (Fig. S9) . (50, 75 , and 100) were tested in lysis buffer (1.2% SDS and pH 11.0) at 20 HAU/mL of H5N3 virus.
TextSentencer_T136 21995-22146 Sentence denotes 50-fold diluted QD650 conjugate to detect H5 HA was mixed with 50-, 70-, and 100-fold diluted QD580 conjugates to detect influenza A NP (A, B, and C) .
TextSentencer_T136 21995-22146 Sentence denotes 50-fold diluted QD650 conjugate to detect H5 HA was mixed with 50-, 70-, and 100-fold diluted QD580 conjugates to detect influenza A NP (A, B, and C) .
TextSentencer_T137 22147-22297 Sentence denotes 70-fold diluted QD650 conjugate to detect H5 HA was mixed with 50-, 70-, and 100-fold diluted QD580 conjugate to detect influenza A NP (D, E, and F) .
TextSentencer_T137 22147-22297 Sentence denotes 70-fold diluted QD650 conjugate to detect H5 HA was mixed with 50-, 70-, and 100-fold diluted QD580 conjugate to detect influenza A NP (D, E, and F) .
TextSentencer_T138 22298-22453 Sentence denotes 100-fold diluted QD650 conjugate to detect H5 HA was mixed with 50-, 70-, and 100-fold diluted QD580 conjugate to detect influenza A NP with (G, H, and I).
TextSentencer_T138 22298-22453 Sentence denotes 100-fold diluted QD650 conjugate to detect H5 HA was mixed with 50-, 70-, and 100-fold diluted QD580 conjugate to detect influenza A NP with (G, H, and I).
TextSentencer_T139 22454-22498 Sentence denotes All data (n = 3) are shown as the mean ± SD.
TextSentencer_T139 22454-22498 Sentence denotes All data (n = 3) are shown as the mean ± SD.
TextSentencer_T140 22499-22805 Sentence denotes F 580-A, TL2/CL of influenza A taken with the 580 nm emission filter; F 580-H5, TL1/CL of H5 taken with the 580 nm emission filter; F 650-A, TL2/CL of influenza A taken with the 650 nm emission filter; F 650-H5, TL1/CL of H5 taken with the 650 nm emission filter. *, P < 0.05; **, P < 0.01; ***, P < 0.001.
TextSentencer_T140 22499-22805 Sentence denotes F 580-A, TL2/CL of influenza A taken with the 580 nm emission filter; F 580-H5, TL1/CL of H5 taken with the 580 nm emission filter; F 650-A, TL2/CL of influenza A taken with the 650 nm emission filter; F 650-H5, TL1/CL of H5 taken with the 650 nm emission filter. *, P < 0.05; **, P < 0.01; ***, P < 0.001.
TextSentencer_T141 22807-22904 Sentence denotes A significant advantage of QDs is that they can be designed to possess distinct emission spectra.
TextSentencer_T141 22807-22904 Sentence denotes A significant advantage of QDs is that they can be designed to possess distinct emission spectra.
TextSentencer_T142 22905-23092 Sentence denotes Furthermore, the emission filter eliminates the fluorescent signal of the other QD, because it absorbs only the wavelengths emitted by each QD and blocks all external fluorescent signals.
TextSentencer_T142 22905-23092 Sentence denotes Furthermore, the emission filter eliminates the fluorescent signal of the other QD, because it absorbs only the wavelengths emitted by each QD and blocks all external fluorescent signals.
TextSentencer_T143 23093-23371 Sentence denotes In this study, the range of optically leaky fluorescent signal derived from the counterpart QD was examined at each emission filter, and the random binding of the antibody conjugate to counterpart test line was analyzed using relatively high titer of virus (up to 1,280 HAU/mL).
TextSentencer_T143 23093-23371 Sentence denotes In this study, the range of optically leaky fluorescent signal derived from the counterpart QD was examined at each emission filter, and the random binding of the antibody conjugate to counterpart test line was analyzed using relatively high titer of virus (up to 1,280 HAU/mL).
TextSentencer_T144 23372-23547 Sentence denotes To replicate the multiplex QD condition of SRDFDS, QD-conjugated mouse IgG (mIgG) was mixed with QD580-conjugated anti-influenza A (7307) or QD650-conjugated anti-H5 HA (2H2).
TextSentencer_T144 23372-23547 Sentence denotes To replicate the multiplex QD condition of SRDFDS, QD-conjugated mouse IgG (mIgG) was mixed with QD580-conjugated anti-influenza A (7307) or QD650-conjugated anti-H5 HA (2H2).
TextSentencer_T145 23548-23681 Sentence denotes The mixture of QD580-conjugated influenza A NP antibody (7307) and QD650-conjugated mIgG was anticipated to be detected at F580 only.
TextSentencer_T145 23548-23681 Sentence denotes The mixture of QD580-conjugated influenza A NP antibody (7307) and QD650-conjugated mIgG was anticipated to be detected at F580 only.
TextSentencer_T146 23682-24071 Sentence denotes However, the TL/CL value derived from this mixture was significantly different at 320 HAU/mL (P <0.01) and 1,280 HAU/mL (P <0.001) for detecting influenza A at F650 implying that the F650 could not eliminate the leaky fluorescent signal derived from the mixture and that the viral concentration of 320-1,280 HAU/mL may also have influenced the cross signaling of QD580 at F650 (Fig. 7A) .
TextSentencer_T146 23682-24071 Sentence denotes However, the TL/CL value derived from this mixture was significantly different at 320 HAU/mL (P <0.01) and 1,280 HAU/mL (P <0.001) for detecting influenza A at F650 implying that the F650 could not eliminate the leaky fluorescent signal derived from the mixture and that the viral concentration of 320-1,280 HAU/mL may also have influenced the cross signaling of QD580 at F650 (Fig. 7A) .
TextSentencer_T147 24072-24266 Sentence denotes Also, QD580-conjugated anti-influenza A NP (7307) antibody did not bind to the test line of H5 HA even at 1,280 HAU/mL, indicating that it did not possess any cross-reactivity with the H5 HA TL.
TextSentencer_T147 24072-24266 Sentence denotes Also, QD580-conjugated anti-influenza A NP (7307) antibody did not bind to the test line of H5 HA even at 1,280 HAU/mL, indicating that it did not possess any cross-reactivity with the H5 HA TL.
TextSentencer_T148 24267-24524 Sentence denotes In the mixture of the QD650-conjugated anti-H5 HA (2H2) and QD580-conjugated mIgG, we found no leaky fluorescent signals in the H5 HA TL at F580 even though TL/CL of H5 HA reached 50, indicating that the F580 totally eliminated the QD650 fluorescent signal.
TextSentencer_T148 24267-24524 Sentence denotes In the mixture of the QD650-conjugated anti-H5 HA (2H2) and QD580-conjugated mIgG, we found no leaky fluorescent signals in the H5 HA TL at F580 even though TL/CL of H5 HA reached 50, indicating that the F580 totally eliminated the QD650 fluorescent signal.
TextSentencer_T149 24525-24714 Sentence denotes However, the same mixture showed significant cross-reactivity (P <0.01) by QD650 conjugate at test line of influenza A NP in the presence of 320 HAU/mL of the H5N3 virus at F650 (Fig. 7B) .
TextSentencer_T149 24525-24714 Sentence denotes However, the same mixture showed significant cross-reactivity (P <0.01) by QD650 conjugate at test line of influenza A NP in the presence of 320 HAU/mL of the H5N3 virus at F650 (Fig. 7B) .
TextSentencer_T150 24715-24907 Sentence denotes This result implied that if the diagnostic result of H5 had a TL/CL value of <10 at ~160 HAU/mL, there was no cross-reaction between QD650-conjugated anti-H5 HA (2H2) and TL of influenza A NP.
TextSentencer_T150 24715-24907 Sentence denotes This result implied that if the diagnostic result of H5 had a TL/CL value of <10 at ~160 HAU/mL, there was no cross-reaction between QD650-conjugated anti-H5 HA (2H2) and TL of influenza A NP.
TextSentencer_T151 24908-25101 Sentence denotes However, at a concentration of >320 HAU/mL with a TL/CL value of >10 in the anti-influenza A NP TL, QD650-conjugated anti-H5 HA appeared to bind to both TLs of the influenza A NP and the H5 HA.
TextSentencer_T151 24908-25101 Sentence denotes However, at a concentration of >320 HAU/mL with a TL/CL value of >10 in the anti-influenza A NP TL, QD650-conjugated anti-H5 HA appeared to bind to both TLs of the influenza A NP and the H5 HA.
TextSentencer_T152 25102-25150 Sentence denotes The raw FICT results are presented in Fig. S10 .
TextSentencer_T152 25102-25150 Sentence denotes The raw FICT results are presented in Fig. S10 .
TextSentencer_T153 25151-25311 Sentence denotes H5 subtype-specificity of SRDFDS was confirmed by testing the H1N1 and H7N1 viruses in specimens at a titer of 100 and 320 HAU/mL using multiplex QD conjugates.
TextSentencer_T153 25151-25311 Sentence denotes H5 subtype-specificity of SRDFDS was confirmed by testing the H1N1 and H7N1 viruses in specimens at a titer of 100 and 320 HAU/mL using multiplex QD conjugates.
TextSentencer_T154 25312-25427 Sentence denotes These samples only showed positive reactions for the influenza A TL but not the H5 HA (P <0.001) (Fig. 7C and 7D) .
TextSentencer_T154 25312-25427 Sentence denotes These samples only showed positive reactions for the influenza A TL but not the H5 HA (P <0.001) (Fig. 7C and 7D) .
TextSentencer_T155 25428-25476 Sentence denotes The raw FICT results are presented in Fig. S11 .
TextSentencer_T155 25428-25476 Sentence denotes The raw FICT results are presented in Fig. S11 .
TextSentencer_T156 25478-25719 Sentence denotes For actual measurements with clinical samples, specimens from 14 patients confirmed to be infected with H5N1 (clade 2.3.4), listed as WHO case numbers, were tested at the National Institute of Hygiene and Epidemiology (NIHE), Hanoi, Vietnam.
TextSentencer_T156 25478-25719 Sentence denotes For actual measurements with clinical samples, specimens from 14 patients confirmed to be infected with H5N1 (clade 2.3.4), listed as WHO case numbers, were tested at the National Institute of Hygiene and Epidemiology (NIHE), Hanoi, Vietnam.
TextSentencer_T157 25720-25808 Sentence denotes The detailed characterization of patients has been described in a previous report [13] .
TextSentencer_T157 25720-25808 Sentence denotes The detailed characterization of patients has been described in a previous report [13] .
TextSentencer_T158 25809-25909 Sentence denotes Patients from # 16 (VN 100) to # 29 (VN 119) were used in our analysis for the evaluation of SRDFDS.
TextSentencer_T158 25809-25909 Sentence denotes Patients from # 16 (VN 100) to # 29 (VN 119) were used in our analysis for the evaluation of SRDFDS.
TextSentencer_T159 25910-26066 Sentence denotes These H5N1-confirmed patients (four females, nine males, and one unknown) with the mean age of 18.5 years (range, 2-40) were enrolled between 2007 and 2010.
TextSentencer_T159 25910-26066 Sentence denotes These H5N1-confirmed patients (four females, nine males, and one unknown) with the mean age of 18.5 years (range, 2-40) were enrolled between 2007 and 2010.
TextSentencer_T160 26067-26141 Sentence denotes Sample collection was performed within a week of the onset of the illness.
TextSentencer_T160 26067-26141 Sentence denotes Sample collection was performed within a week of the onset of the illness.
TextSentencer_T161 26142-26258 Sentence denotes Among the 14 cases, 7 patients died within two weeks of the onset of the illness, showing 50% mortality ( Table 2) .
TextSentencer_T161 26142-26258 Sentence denotes Among the 14 cases, 7 patients died within two weeks of the onset of the illness, showing 50% mortality ( Table 2) .
TextSentencer_T162 26259-26561 Sentence denotes For binary diagnostic decision, the TL/CL threshold cut-off value for H5N1 was determined to be 0.77 for influenza A and 0.25 for H5 HA from receiver-operating characteristic (ROC) curve analysis; all TL/CL values were normalized based on this cut-off value (positive if TL/CL > 1, negative otherwise).
TextSentencer_T162 26259-26561 Sentence denotes For binary diagnostic decision, the TL/CL threshold cut-off value for H5N1 was determined to be 0.77 for influenza A and 0.25 for H5 HA from receiver-operating characteristic (ROC) curve analysis; all TL/CL values were normalized based on this cut-off value (positive if TL/CL > 1, negative otherwise).
TextSentencer_T163 26562-26671 Sentence denotes The ROC curve analysis resulted in an area under the curve (AUC) value of 0.96 (95% confidence interval [CI]:
TextSentencer_T163 26562-26671 Sentence denotes The ROC curve analysis resulted in an area under the curve (AUC) value of 0.96 (95% confidence interval [CI]:
TextSentencer_T164 26672-26741 Sentence denotes 0.89-1.033) for the diagnosis of influenza A (P < 0.0001) (Fig. 8A) .
TextSentencer_T164 26672-26741 Sentence denotes 0.89-1.033) for the diagnosis of influenza A (P < 0.0001) (Fig. 8A) .
TextSentencer_T165 26742-26795 Sentence denotes The diagnosis of H5 HA showed an AUC of 0.89 (95% CI:
TextSentencer_T165 26742-26795 Sentence denotes The diagnosis of H5 HA showed an AUC of 0.89 (95% CI:
TextSentencer_T166 26796-26930 Sentence denotes 0.76-1.008; P < 0.0001) (Fig. 8B) , indicating that SRDFDS had high accuracy to predict H5N1 infection for both influenza A and H5 HA.
TextSentencer_T166 26796-26930 Sentence denotes 0.76-1.008; P < 0.0001) (Fig. 8B) , indicating that SRDFDS had high accuracy to predict H5N1 infection for both influenza A and H5 HA.
TextSentencer_T167 26931-27111 Sentence denotes Of the H5N1 positive 14 patients, only one case showed a false-negative TL/CL value while the other cases showed above the threshold TL/CL values for influenza A detection at F580.
TextSentencer_T167 26931-27111 Sentence denotes Of the H5N1 positive 14 patients, only one case showed a false-negative TL/CL value while the other cases showed above the threshold TL/CL values for influenza A detection at F580.
TextSentencer_T168 27112-27210 Sentence denotes The non-infected control group (n = 38) showed no false-positive cases for influenza A (Fig. 8C) .
TextSentencer_T168 27112-27210 Sentence denotes The non-infected control group (n = 38) showed no false-positive cases for influenza A (Fig. 8C) .
TextSentencer_T169 27211-27370 Sentence denotes When detecting H5 HA in the patients, SRDFDS showed eleven positive cases and three false negative cases based on the threshold TL/CL value at F650 (Fig. 8D) .
TextSentencer_T169 27211-27370 Sentence denotes When detecting H5 HA in the patients, SRDFDS showed eleven positive cases and three false negative cases based on the threshold TL/CL value at F650 (Fig. 8D) .
TextSentencer_T170 27371-27493 Sentence denotes Five cases with a higher TL/CL value than the other cases showed cross-reactivity at different emission filter (Fig. 8B) .
TextSentencer_T170 27371-27493 Sentence denotes Five cases with a higher TL/CL value than the other cases showed cross-reactivity at different emission filter (Fig. 8B) .
TextSentencer_T171 27494-27609 Sentence denotes Four out of five cases with a positive TL/CL at F580 were also strongly positive at F650 for influenza A detection.
TextSentencer_T171 27494-27609 Sentence denotes Four out of five cases with a positive TL/CL at F580 were also strongly positive at F650 for influenza A detection.
TextSentencer_T172 27610-27694 Sentence denotes Detailed results of the TL/CL values from SRDFDS and RDT are presented in Table 3 .
TextSentencer_T172 27610-27694 Sentence denotes Detailed results of the TL/CL values from SRDFDS and RDT are presented in Table 3 .
TextSentencer_T173 27695-27711 Sentence denotes H5 subtype (B) .
TextSentencer_T173 27695-27711 Sentence denotes H5 subtype (B) .
TextSentencer_T174 27712-27798 Sentence denotes The accuracy of the SRDFDS was determined based on the area under the ROC curve (AUC).
TextSentencer_T174 27712-27798 Sentence denotes The accuracy of the SRDFDS was determined based on the area under the ROC curve (AUC).
TextSentencer_T175 27799-27899 Sentence denotes The cut-off value used for the detection of H5N1 was applied to determine the presence of the virus.
TextSentencer_T175 27799-27899 Sentence denotes The cut-off value used for the detection of H5N1 was applied to determine the presence of the virus.
TextSentencer_T176 27900-28151 Sentence denotes All fluorescence levels of TL/CL against negative control group or positive patients were plotted for prediction of infection by influenza A and H5 subtype using two different emission filters for 580 nm (F580) (C) and 650 nm (F650) (D), respectively.
TextSentencer_T176 27900-28151 Sentence denotes All fluorescence levels of TL/CL against negative control group or positive patients were plotted for prediction of infection by influenza A and H5 subtype using two different emission filters for 580 nm (F580) (C) and 650 nm (F650) (D), respectively.
TextSentencer_T177 28152-28335 Sentence denotes Among the negative control patients, only one case of a metapneumovirus (MPV)-infected patient showed a false-positive for H5 HA detection with higher TL/CL values than the threshold.
TextSentencer_T177 28152-28335 Sentence denotes Among the negative control patients, only one case of a metapneumovirus (MPV)-infected patient showed a false-positive for H5 HA detection with higher TL/CL values than the threshold.
TextSentencer_T178 28336-28432 Sentence denotes As seen in Table 3 38) showed no false-positive cases for the detection of influenza A or H5 HA.
TextSentencer_T178 28336-28432 Sentence denotes As seen in Table 3 38) showed no false-positive cases for the detection of influenza A or H5 HA.
TextSentencer_T179 28433-28476 Sentence denotes All results from RDT are presented in Figs.
TextSentencer_T179 28433-28476 Sentence denotes All results from RDT are presented in Figs.
TextSentencer_T180 28477-28489 Sentence denotes S14 and S15.
TextSentencer_T180 28477-28489 Sentence denotes S14 and S15.
TextSentencer_T181 28490-28652 Sentence denotes Based on these results, the H5N1-specific SRDFDS we developed had superior specificity and sensitivity for the detection of H5N1 HPAI virus strains (clade 2.3.4).
TextSentencer_T181 28490-28652 Sentence denotes Based on these results, the H5N1-specific SRDFDS we developed had superior specificity and sensitivity for the detection of H5N1 HPAI virus strains (clade 2.3.4).
TextSentencer_T182 28653-28807 Sentence denotes The reactivity of mAbs to other H5N1 (clade 1) was also confirmed, indicating the capability of SRDFDS to detect multiple clades of H5N1 HPAI (Fig. S16 ).
TextSentencer_T182 28653-28807 Sentence denotes The reactivity of mAbs to other H5N1 (clade 1) was also confirmed, indicating the capability of SRDFDS to detect multiple clades of H5N1 HPAI (Fig. S16 ).
TextSentencer_T183 28808-28991 Sentence denotes Since November 2003, WHO reported more than 700 human cases of Asian HPAI H5N1 viruses mostly from fifteen countries across Asia, Africa, the Pacific, Europe, and the Near East [42] .
TextSentencer_T183 28808-28991 Sentence denotes Since November 2003, WHO reported more than 700 human cases of Asian HPAI H5N1 viruses mostly from fifteen countries across Asia, Africa, the Pacific, Europe, and the Near East [42] .
TextSentencer_T184 28992-29167 Sentence denotes Due to high crossreactivity and antigenic variability, H5-subtypespecific rapid diagnostic systems have been developed and tested mainly in wild birds and not in humans [43] .
TextSentencer_T184 28992-29167 Sentence denotes Due to high crossreactivity and antigenic variability, H5-subtypespecific rapid diagnostic systems have been developed and tested mainly in wild birds and not in humans [43] .
TextSentencer_T185 29168-29280 Sentence denotes Therefore, there is an urgent need to develop more sensitive and H5 subtype-specific on-site monitoring systems.
TextSentencer_T185 29168-29280 Sentence denotes Therefore, there is an urgent need to develop more sensitive and H5 subtype-specific on-site monitoring systems.
TextSentencer_T186 29281-29366 Sentence denotes However, H5-subtypespecific rapid diagnostic systems have only been rarely developed.
TextSentencer_T186 29281-29366 Sentence denotes However, H5-subtypespecific rapid diagnostic systems have only been rarely developed.
TextSentencer_T187 29367-29518 Sentence denotes QD-based electrochemical assays have been extensively explored because of their optical properties, tunability, and potential for multiplexing in POCT.
TextSentencer_T187 29367-29518 Sentence denotes QD-based electrochemical assays have been extensively explored because of their optical properties, tunability, and potential for multiplexing in POCT.
TextSentencer_T188 29519-29826 Sentence denotes So far, preventing nonspecific reactions in QDs and increasing solubility have been major challenges because the use of high-temperature organometallic synthetic procedures produces monodisperse nanocrystals with a passivated surface containing a monolayer of hydrophobic surfactant ligands [44] [45] [46] .
TextSentencer_T188 29519-29826 Sentence denotes So far, preventing nonspecific reactions in QDs and increasing solubility have been major challenges because the use of high-temperature organometallic synthetic procedures produces monodisperse nanocrystals with a passivated surface containing a monolayer of hydrophobic surfactant ligands [44] [45] [46] .
TextSentencer_T189 29827-30044 Sentence denotes QDs are insolubly monodispersed with a very wide range of possible emission spectra ranging from ultraviolet to near-infrared that can be controlled by simply changing their size, composition, and structure [36, 37] .
TextSentencer_T189 29827-30044 Sentence denotes QDs are insolubly monodispersed with a very wide range of possible emission spectra ranging from ultraviolet to near-infrared that can be controlled by simply changing their size, composition, and structure [36, 37] .
TextSentencer_T190 30045-30272 Sentence denotes For water compatibility and bioconjugation, QDs are covered with a hydrophilic shell which is a popular method to solubilize QDs in aqueous media and improve their signal intensity through encapsulation with polymers [31, 32] .
TextSentencer_T190 30045-30272 Sentence denotes For water compatibility and bioconjugation, QDs are covered with a hydrophilic shell which is a popular method to solubilize QDs in aqueous media and improve their signal intensity through encapsulation with polymers [31, 32] .
TextSentencer_T191 30273-30476 Sentence denotes To apply QDs to POCT, encapsulation of QDs with a polymer was routinely used to amplify the fluorescent signal, and few trials have attempted to use polymers on QD surface for POCT systems [31, 32, 47] .
TextSentencer_T191 30273-30476 Sentence denotes To apply QDs to POCT, encapsulation of QDs with a polymer was routinely used to amplify the fluorescent signal, and few trials have attempted to use polymers on QD surface for POCT systems [31, 32, 47] .
TextSentencer_T192 30477-30583 Sentence denotes This may reflect the weakness of the fluorescent signal of QDs making it difficult to be captured by POCT.
TextSentencer_T192 30477-30583 Sentence denotes This may reflect the weakness of the fluorescent signal of QDs making it difficult to be captured by POCT.
TextSentencer_T193 30584-30727 Sentence denotes An additional method of encapsulating many QDs in one particle was suggested to amplify the fluorescent signal from the QD nanoparticles [32] .
TextSentencer_T193 30584-30727 Sentence denotes An additional method of encapsulating many QDs in one particle was suggested to amplify the fluorescent signal from the QD nanoparticles [32] .
TextSentencer_T194 30728-30881 Sentence denotes Currently, a rapid dual POCT system has been developed that detects two different pathogen species rather than two antigen targets in one pathogen [33] .
TextSentencer_T194 30728-30881 Sentence denotes Currently, a rapid dual POCT system has been developed that detects two different pathogen species rather than two antigen targets in one pathogen [33] .
TextSentencer_T195 30882-31190 Sentence denotes However, simultaneous detection of two antigens of one pathogen, HPAI H5N1, would strengthen the primary screening of POCT and inform clinicians of the H5-infection potential, although the patient may test negative due to the relatively small amount of H5-specific antigen compared to influenza A NP antigen.
TextSentencer_T195 30882-31190 Sentence denotes However, simultaneous detection of two antigens of one pathogen, HPAI H5N1, would strengthen the primary screening of POCT and inform clinicians of the H5-infection potential, although the patient may test negative due to the relatively small amount of H5-specific antigen compared to influenza A NP antigen.
TextSentencer_T196 31191-31370 Sentence denotes To overcome this limitation, in this study, we developed a novel method using CdSe/CdS/ZnS QDs for POCT and evaluated the clinical feasibility of this multiplex diagnostic system.
TextSentencer_T196 31191-31370 Sentence denotes To overcome this limitation, in this study, we developed a novel method using CdSe/CdS/ZnS QDs for POCT and evaluated the clinical feasibility of this multiplex diagnostic system.
TextSentencer_T197 31371-31568 Sentence denotes We also employed a unique method for coating QDs on the surface of a latex increasing the sensitivity of the fluorescent signal, thus improving the sensitivity of the POCT system (Fig. 1A and 1E) .
TextSentencer_T197 31371-31568 Sentence denotes We also employed a unique method for coating QDs on the surface of a latex increasing the sensitivity of the fluorescent signal, thus improving the sensitivity of the POCT system (Fig. 1A and 1E) .
TextSentencer_T198 31569-31704 Sentence denotes In order to increase the accuracy of POCT, each TL value on the lateral flow strip needs to be normalized against that of the CL [30] .
TextSentencer_T198 31569-31704 Sentence denotes In order to increase the accuracy of POCT, each TL value on the lateral flow strip needs to be normalized against that of the CL [30] .
TextSentencer_T199 31705-31779 Sentence denotes In a previous study, a single QD was used to detect two different viruses.
TextSentencer_T199 31705-31779 Sentence denotes In a previous study, a single QD was used to detect two different viruses.
TextSentencer_T200 31780-32066 Sentence denotes Most of the multiplex QD-labeled strips, which possessed an extremely high sensitivity (0.016 HAU of H5 and 0.25 HAU of influenza A as a limit of detection), were used without accurate normalization of the CL resulting in qualitative rather than quantitative detection of targets [33] .
TextSentencer_T200 31780-32066 Sentence denotes Most of the multiplex QD-labeled strips, which possessed an extremely high sensitivity (0.016 HAU of H5 and 0.25 HAU of influenza A as a limit of detection), were used without accurate normalization of the CL resulting in qualitative rather than quantitative detection of targets [33] .
TextSentencer_T201 32067-32242 Sentence denotes Other groups have applied a single QD for detecting only one target antigen of one viral particle rather than for simultaneous targeting of two antigens of one viral particle.
TextSentencer_T201 32067-32242 Sentence denotes Other groups have applied a single QD for detecting only one target antigen of one viral particle rather than for simultaneous targeting of two antigens of one viral particle.
TextSentencer_T202 32243-32493 Sentence denotes Chen and colleagues studied fluorescence-linked immunosorbent assay (FLISA) with chicken samples and Cui et al. examined influenza virus using an electron microscope for imaging; however, none of these systems was a rapid diagnostic system [35, 48] .
TextSentencer_T202 32243-32493 Sentence denotes Chen and colleagues studied fluorescence-linked immunosorbent assay (FLISA) with chicken samples and Cui et al. examined influenza virus using an electron microscope for imaging; however, none of these systems was a rapid diagnostic system [35, 48] .
TextSentencer_T203 32494-32644 Sentence denotes Li's group developed a QD-linked rapid diagnostic system, which needed additional HCL and Br2 treatment steps to measure fluorescence intensity [49] .
TextSentencer_T203 32494-32644 Sentence denotes Li's group developed a QD-linked rapid diagnostic system, which needed additional HCL and Br2 treatment steps to measure fluorescence intensity [49] .
TextSentencer_T204 32645-32798 Sentence denotes In contrast, our assay provides a rapid system for detecting two antigens of one virus simultaneously resulting in the reliability of the screening data.
TextSentencer_T204 32645-32798 Sentence denotes In contrast, our assay provides a rapid system for detecting two antigens of one virus simultaneously resulting in the reliability of the screening data.
TextSentencer_T205 32799-32923 Sentence denotes Two different QDs and emission filters were used for targeting two different components of the same virus on a single strip.
TextSentencer_T205 32799-32923 Sentence denotes Two different QDs and emission filters were used for targeting two different components of the same virus on a single strip.
TextSentencer_T206 32924-33049 Sentence denotes All components of the viral antigens needed to be separated from the virion to prevent cross-reactions on the counterpart TL.
TextSentencer_T206 32924-33049 Sentence denotes All components of the viral antigens needed to be separated from the virion to prevent cross-reactions on the counterpart TL.
TextSentencer_T207 33050-33239 Sentence denotes In our study, the bioconjugate of latex, QDs, and mAb was used for the simultaneous identification of influenza A and H5 subtype with no cross-reactivity at a concentration of 5-160 HAU/mL.
TextSentencer_T207 33050-33239 Sentence denotes In our study, the bioconjugate of latex, QDs, and mAb was used for the simultaneous identification of influenza A and H5 subtype with no cross-reactivity at a concentration of 5-160 HAU/mL.
TextSentencer_T208 33240-33388 Sentence denotes This assay showed a good linear regression at a TL/CL range of 1-10 (Fig. 6) , which corresponds to the TL/CL ranges in clinical patients (Fig. 8) .
TextSentencer_T208 33240-33388 Sentence denotes This assay showed a good linear regression at a TL/CL range of 1-10 (Fig. 6) , which corresponds to the TL/CL ranges in clinical patients (Fig. 8) .
TextSentencer_T209 33389-33532 Sentence denotes As reported by de Jong et al. a fatal case had H5N1 viral loads of about 5.8 log10 cDNA copies (nose) and 7.5 log10 cDNA copies (throat) [50] .
TextSentencer_T209 33389-33532 Sentence denotes As reported by de Jong et al. a fatal case had H5N1 viral loads of about 5.8 log10 cDNA copies (nose) and 7.5 log10 cDNA copies (throat) [50] .
TextSentencer_T210 33533-33724 Sentence denotes The rRT-PCR in our study of 5.9 log 10 cDNA corresponded to the limit of detection of our assay for H5 HA and 5 -6.5 log 10 cDNA copies corresponded to the quantitative range of 5-160 HAU/mL.
TextSentencer_T210 33533-33724 Sentence denotes The rRT-PCR in our study of 5.9 log 10 cDNA corresponded to the limit of detection of our assay for H5 HA and 5 -6.5 log 10 cDNA copies corresponded to the quantitative range of 5-160 HAU/mL.
TextSentencer_T211 33725-33848 Sentence denotes Taken together, we believe that our limit of detection and quantitative range enabled detection of a fatal H5N1 human case.
TextSentencer_T211 33725-33848 Sentence denotes Taken together, we believe that our limit of detection and quantitative range enabled detection of a fatal H5N1 human case.
TextSentencer_T212 33849-33994 Sentence denotes We have provided evidence by diluting HPAI H5N1 virus in addition to H5N3, which showed a good linear regression in this TL/CL range (Fig. S17) .
TextSentencer_T212 33849-33994 Sentence denotes We have provided evidence by diluting HPAI H5N1 virus in addition to H5N3, which showed a good linear regression in this TL/CL range (Fig. S17) .
TextSentencer_T213 33995-34169 Sentence denotes As Fig. 7 shows, the cross-reactivity of influenza A TL obtained by F650 nm at 160 HAU/mL was not derived from the random binding of QD650 conjugate to influenza A test line.
TextSentencer_T213 33995-34169 Sentence denotes As Fig. 7 shows, the cross-reactivity of influenza A TL obtained by F650 nm at 160 HAU/mL was not derived from the random binding of QD650 conjugate to influenza A test line.
TextSentencer_T214 34170-34435 Sentence denotes Except for one patient (VN 102) (TL/CL;1.31), four patients (VN 100, VN 105, VN 106, and VN 109) , who had TL/CL value of > 3 against influenza A detection, were positive at F650 and were not due to the random binding of QD650 conjugate but simple optical leaking.
TextSentencer_T214 34170-34435 Sentence denotes Except for one patient (VN 102) (TL/CL;1.31), four patients (VN 100, VN 105, VN 106, and VN 109) , who had TL/CL value of > 3 against influenza A detection, were positive at F650 and were not due to the random binding of QD650 conjugate but simple optical leaking.
TextSentencer_T215 34436-34621 Sentence denotes As this optical leaking did not interfere with TL1/CL or TL2/CL results in patients, clinical readouts to diagnose influenza A and H5 HA measured by SRDFDS could be considered reliable.
TextSentencer_T215 34436-34621 Sentence denotes As this optical leaking did not interfere with TL1/CL or TL2/CL results in patients, clinical readouts to diagnose influenza A and H5 HA measured by SRDFDS could be considered reliable.
TextSentencer_T216 34622-34865 Sentence denotes Our system was limited by random binding of QD650 conjugate at influenza A TL at an extremely high titer, such as 320-1200 HAU/mL, due to the use of a less than perfect lysis buffer, where the fluorescence signal of QD580 was captured by F650.
TextSentencer_T216 34622-34865 Sentence denotes Our system was limited by random binding of QD650 conjugate at influenza A TL at an extremely high titer, such as 320-1200 HAU/mL, due to the use of a less than perfect lysis buffer, where the fluorescence signal of QD580 was captured by F650.
TextSentencer_T217 34866-35196 Sentence denotes In contrast, the F580 excluded the fluorescent signal from QD650 conjugates completely while QD650 conjugates targeting H5 HA were cross-reactive to the TL of influenza A NP at concentrations greater than 320 HAU/mL, indicating that the NP-and HA antigens of single virus particle were not completely separated at this high titer.
TextSentencer_T217 34866-35196 Sentence denotes In contrast, the F580 excluded the fluorescent signal from QD650 conjugates completely while QD650 conjugates targeting H5 HA were cross-reactive to the TL of influenza A NP at concentrations greater than 320 HAU/mL, indicating that the NP-and HA antigens of single virus particle were not completely separated at this high titer.
TextSentencer_T218 35197-35422 Sentence denotes For influenza A diagnosis, an isothermal nucleic acid amplification-based integrated system, Alere i influenza A&B test, had an excellent performance showing a sensitivity of 87.2-93.8% and specificity of 62.5-100% [51, 52] .
TextSentencer_T218 35197-35422 Sentence denotes For influenza A diagnosis, an isothermal nucleic acid amplification-based integrated system, Alere i influenza A&B test, had an excellent performance showing a sensitivity of 87.2-93.8% and specificity of 62.5-100% [51, 52] .
TextSentencer_T219 35423-35499 Sentence denotes Moreover, an antigen detection kit showed a sensitivity of 60-80% [53, 54] .
TextSentencer_T219 35423-35499 Sentence denotes Moreover, an antigen detection kit showed a sensitivity of 60-80% [53, 54] .
TextSentencer_T220 35500-35681 Sentence denotes Therefore, our result is comparable to that of the isothermal nucleic acid amplification-based integrated system and could be more accurate than that of the rapid diagnostic system.
TextSentencer_T220 35500-35681 Sentence denotes Therefore, our result is comparable to that of the isothermal nucleic acid amplification-based integrated system and could be more accurate than that of the rapid diagnostic system.
TextSentencer_T221 35682-35776 Sentence denotes However, no commercialized H5 subtype-specific rapid diagnostic system is currently available.
TextSentencer_T221 35682-35776 Sentence denotes However, no commercialized H5 subtype-specific rapid diagnostic system is currently available.
TextSentencer_T222 35777-35975 Sentence denotes Here, we have developed an assay that provides a reliable primary screening result to clinicians by furnishing dual information for highly pathogenic H5N1 virus-infected patients with 50% mortality.
TextSentencer_T222 35777-35975 Sentence denotes Here, we have developed an assay that provides a reliable primary screening result to clinicians by furnishing dual information for highly pathogenic H5N1 virus-infected patients with 50% mortality.
TextSentencer_T223 35976-36159 Sentence denotes As laboratory immunoassay, we conducted sandwich fluorescence-linked immunosorbent assay (FLISA)with the QD conjugates and found that SRDFDS was more sensitive than FLISA (Fig. S18) .
TextSentencer_T223 35976-36159 Sentence denotes As laboratory immunoassay, we conducted sandwich fluorescence-linked immunosorbent assay (FLISA)with the QD conjugates and found that SRDFDS was more sensitive than FLISA (Fig. S18) .
TextSentencer_T224 36160-36376 Sentence denotes Furthermore, the lowest virus titer of a commercial RDT (influenza A) was 80 HAU/mL, indicating that our SRDFDS improved the performance to recognize the same virus by 8 folds over the commercial RDT kit (Fig. S19) .
TextSentencer_T224 36160-36376 Sentence denotes Furthermore, the lowest virus titer of a commercial RDT (influenza A) was 80 HAU/mL, indicating that our SRDFDS improved the performance to recognize the same virus by 8 folds over the commercial RDT kit (Fig. S19) .
TextSentencer_T225 36377-36551 Sentence denotes To the best of our knowledge, our study is the first to clinically evaluate a rapid kit with the ability to detect influenza A and H5-subtype simultaneously in human samples.
TextSentencer_T225 36377-36551 Sentence denotes To the best of our knowledge, our study is the first to clinically evaluate a rapid kit with the ability to detect influenza A and H5-subtype simultaneously in human samples.
TextSentencer_T226 36552-36766 Sentence denotes This assay was designed to develop an efficient QD-based method with more than 50% QY; we believe that our study could adapt current QDs to the POCT platform based on lateral flow test strips via latex conjugation.
TextSentencer_T226 36552-36766 Sentence denotes This assay was designed to develop an efficient QD-based method with more than 50% QY; we believe that our study could adapt current QDs to the POCT platform based on lateral flow test strips via latex conjugation.
TextSentencer_T227 36767-36975 Sentence denotes In summary, the H5 subtype-specific SRDFDS developed in this study allowed for the accurate identification of influenza A and HA subtype for the rapid detection of H5N1 HPAI viruses in human clinical samples.
TextSentencer_T227 36767-36975 Sentence denotes In summary, the H5 subtype-specific SRDFDS developed in this study allowed for the accurate identification of influenza A and HA subtype for the rapid detection of H5N1 HPAI viruses in human clinical samples.
TextSentencer_T228 36976-37154 Sentence denotes This multiplexing of the QD-based SRDFDS system can lay the foundation for the rapid and simultaneous detection of subtype-specific influenza virus samples in a clinical setting.
TextSentencer_T228 36976-37154 Sentence denotes This multiplexing of the QD-based SRDFDS system can lay the foundation for the rapid and simultaneous detection of subtype-specific influenza virus samples in a clinical setting.
TextSentencer_T229 37155-37231 Sentence denotes Program of the NRF funded by the Korean government, MSIP (2014M3A9E2064699).
TextSentencer_T229 37155-37231 Sentence denotes Program of the NRF funded by the Korean government, MSIP (2014M3A9E2064699).
TextSentencer_T230 37232-37314 Sentence denotes SY and HK designed and optimized all immunoassays under the guidance of HC and HP.
TextSentencer_T230 37232-37314 Sentence denotes SY and HK designed and optimized all immunoassays under the guidance of HC and HP.
TextSentencer_T231 37315-37361 Sentence denotes TD synthesized and characterized quantum dots.
TextSentencer_T231 37315-37361 Sentence denotes TD synthesized and characterized quantum dots.
TextSentencer_T232 37362-37404 Sentence denotes BC, NA, and TT conducted all immunoassays.
TextSentencer_T232 37362-37404 Sentence denotes BC, NA, and TT conducted all immunoassays.
TextSentencer_T233 37405-37453 Sentence denotes NH, HP, LT, and LM conducted the clinical study.
TextSentencer_T233 37405-37453 Sentence denotes NH, HP, LT, and LM conducted the clinical study.
TextSentencer_T234 37454-37496 Sentence denotes YR and KY developed the diagnostic device.
TextSentencer_T234 37454-37496 Sentence denotes YR and KY developed the diagnostic device.
TextSentencer_T235 37497-37547 Sentence denotes HS and CC developed monoclonal antibody and strip.
TextSentencer_T235 37497-37547 Sentence denotes HS and CC developed monoclonal antibody and strip.
TextSentencer_T236 37548-37624 Sentence denotes SY, HK, HC, and HP wrote the paper with input from all other authors. [55] .
TextSentencer_T236 37548-37624 Sentence denotes SY, HK, HC, and HP wrote the paper with input from all other authors. [55] .
TextSentencer_T237 37625-37763 Sentence denotes The TOPO-capped CdSe/CdS/ZnS QD multi-shells were synthesized using the successive ion layer adsorption and reaction (SILAR) method [56] .
TextSentencer_T237 37625-37763 Sentence denotes The TOPO-capped CdSe/CdS/ZnS QD multi-shells were synthesized using the successive ion layer adsorption and reaction (SILAR) method [56] .
TextSentencer_T238 37764-37867 Sentence denotes The CdS and ZnS shells were grown consecutively over a CdSe core at an optimized temperature of 240 °C.
TextSentencer_T238 37764-37867 Sentence denotes The CdS and ZnS shells were grown consecutively over a CdSe core at an optimized temperature of 240 °C.
TextSentencer_T239 37868-38043 Sentence denotes Water-soluble MPA-capped CdSe/CdS/ZnS QDs were prepared by exchanging the TOPO ligand in the synthesized TOPO-capped CdSe/CdS/ZnS QDs with the 3-mercaptopropionic acid ligand.
TextSentencer_T239 37868-38043 Sentence denotes Water-soluble MPA-capped CdSe/CdS/ZnS QDs were prepared by exchanging the TOPO ligand in the synthesized TOPO-capped CdSe/CdS/ZnS QDs with the 3-mercaptopropionic acid ligand.
TextSentencer_T240 38044-38256 Sentence denotes The reaction conditions in this ligand-exchange method were achieved by using tetramethylammonium hydroxide (TMAOH), an organic base, to improve the reactivity of the thiol with the CdSe/CdS/ZnS QD surface [57] .
TextSentencer_T240 38044-38256 Sentence denotes The reaction conditions in this ligand-exchange method were achieved by using tetramethylammonium hydroxide (TMAOH), an organic base, to improve the reactivity of the thiol with the CdSe/CdS/ZnS QD surface [57] .
TextSentencer_T241 38257-38347 Sentence denotes The detailed procedure for the synthesis of QDs is described in the Supplementary Methods.
TextSentencer_T241 38257-38347 Sentence denotes The detailed procedure for the synthesis of QDs is described in the Supplementary Methods.
TextSentencer_T242 38348-38654 Sentence denotes One H5 subtype-specific monoclonal antibody (mAb) (2H2) (detection) was developed by Professor Ho-Joon Shin (Ajou University, Suwon, Korea) and Professor Hyun Park (Wonkwnag University, Iksan, Korea); H5 subtype-specific mAb (1C5) (capture) was donated by Dr. Chom-Kyu Chong (GenBody Inc., Cheonan, Korea).
TextSentencer_T242 38348-38654 Sentence denotes One H5 subtype-specific monoclonal antibody (mAb) (2H2) (detection) was developed by Professor Ho-Joon Shin (Ajou University, Suwon, Korea) and Professor Hyun Park (Wonkwnag University, Iksan, Korea); H5 subtype-specific mAb (1C5) (capture) was donated by Dr. Chom-Kyu Chong (GenBody Inc., Cheonan, Korea).
TextSentencer_T243 38655-38821 Sentence denotes Two monoclonal antibodies, anti-influenza A-7307 (detection) and A-7304 (capture) targeting nucleoprotein (NP), were purchased from Medix Biochemica (Espoo, Finland).
TextSentencer_T243 38655-38821 Sentence denotes Two monoclonal antibodies, anti-influenza A-7307 (detection) and A-7304 (capture) targeting nucleoprotein (NP), were purchased from Medix Biochemica (Espoo, Finland).
TextSentencer_T244 38822-38998 Sentence denotes High-resolution transmission electron microscopy (HR-TEM) was performed using a JEM-2010F electron microscope (JEOL, Tokyo, Japan) with an accelerating voltage of 200 kV [58] .
TextSentencer_T244 38822-38998 Sentence denotes High-resolution transmission electron microscopy (HR-TEM) was performed using a JEM-2010F electron microscope (JEOL, Tokyo, Japan) with an accelerating voltage of 200 kV [58] .
TextSentencer_T245 38999-39134 Sentence denotes For TEM sampling, 0.2% sample solutions were drop-casted onto a carbon-coated copper grid and the solvents were evaporated in a vacuum.
TextSentencer_T245 38999-39134 Sentence denotes For TEM sampling, 0.2% sample solutions were drop-casted onto a carbon-coated copper grid and the solvents were evaporated in a vacuum.
TextSentencer_T246 39135-39210 Sentence denotes UV-Vis absorption spectra were obtained using a SCINCO S-3150 spectrometer.
TextSentencer_T246 39135-39210 Sentence denotes UV-Vis absorption spectra were obtained using a SCINCO S-3150 spectrometer.
TextSentencer_T247 39211-39465 Sentence denotes Photoluminescence (PL) spectroscopy and QY measurements were performed using a He-Cd (Kimmon Electric Co., IK3501R-G, Tokyo, Japan) excitation source at 480 nm, as well as a photodiode array detector (IRY1024, Princeton Instrument Co., Trenton, NJ, USA).
TextSentencer_T247 39211-39465 Sentence denotes Photoluminescence (PL) spectroscopy and QY measurements were performed using a He-Cd (Kimmon Electric Co., IK3501R-G, Tokyo, Japan) excitation source at 480 nm, as well as a photodiode array detector (IRY1024, Princeton Instrument Co., Trenton, NJ, USA).
TextSentencer_T248 39466-39590 Sentence denotes Nuclear magnetic resonance (NMR) data were collected using a superconducting FT-NMR at 300 MHz (Varian, Palo Alto, CA, USA).
TextSentencer_T248 39466-39590 Sentence denotes Nuclear magnetic resonance (NMR) data were collected using a superconducting FT-NMR at 300 MHz (Varian, Palo Alto, CA, USA).
TextSentencer_T249 39591-39648 Sentence denotes Chemical shifts were reported in parts per million (ppm).
TextSentencer_T249 39591-39648 Sentence denotes Chemical shifts were reported in parts per million (ppm).
TextSentencer_T250 39649-39860 Sentence denotes High-resolution X-ray diffraction (XRD) patterns were collected using an X'Pert PRO Multi-Purpose X-ray diffractometer (PANalytical, Almelo, Netherlands) equipped with a Cu Kα source operated at 40 kV and 30 mA.
TextSentencer_T250 39649-39860 Sentence denotes High-resolution X-ray diffraction (XRD) patterns were collected using an X'Pert PRO Multi-Purpose X-ray diffractometer (PANalytical, Almelo, Netherlands) equipped with a Cu Kα source operated at 40 kV and 30 mA.
TextSentencer_T251 39861-39949 Sentence denotes Thermogravimetric analysis (TGA) was conducted using a METTLER TOLEDO SDTA851e under N2.
TextSentencer_T251 39861-39949 Sentence denotes Thermogravimetric analysis (TGA) was conducted using a METTLER TOLEDO SDTA851e under N2.
TextSentencer_T252 39950-40077 Sentence denotes Avian influenza virus H5N3, H7N1, H1N1, and influenza B were obtained from the Centers for Disease Control & Prevention, Korea.
TextSentencer_T252 39950-40077 Sentence denotes Avian influenza virus H5N3, H7N1, H1N1, and influenza B were obtained from the Centers for Disease Control & Prevention, Korea.
TextSentencer_T253 40078-40194 Sentence denotes All viruses were cultured by egg inoculation and titrated by a hemagglutination assay as described previously [12] .
TextSentencer_T253 40078-40194 Sentence denotes All viruses were cultured by egg inoculation and titrated by a hemagglutination assay as described previously [12] .
TextSentencer_T254 40195-40467 Sentence denotes Next, 400 µL succinic anhydride (60 mg/mL) in 0.1 M sodium phosphate was added to completely exchange -NH2 groups to -COOH groups on the surface of the latex-QD complex which was activated by adding 1.3 µL 50 mM EDC and 10 µL 0.5 M Sulfo-NHS in 0.05 M MES buffer (pH 6.1).
TextSentencer_T254 40195-40467 Sentence denotes Next, 400 µL succinic anhydride (60 mg/mL) in 0.1 M sodium phosphate was added to completely exchange -NH2 groups to -COOH groups on the surface of the latex-QD complex which was activated by adding 1.3 µL 50 mM EDC and 10 µL 0.5 M Sulfo-NHS in 0.05 M MES buffer (pH 6.1).
TextSentencer_T255 40468-40607 Sentence denotes Activated latex-QDs were then mixed with 75 µL antibody (1 mg/mL) in 1 mL 0.05 M MES buffer (pH 6.1) and allowed to react for 6 h at 25 °C.
TextSentencer_T255 40468-40607 Sentence denotes Activated latex-QDs were then mixed with 75 µL antibody (1 mg/mL) in 1 mL 0.05 M MES buffer (pH 6.1) and allowed to react for 6 h at 25 °C.
TextSentencer_T256 40608-40827 Sentence denotes After centrifugation at 27,237 × g for 5 min, the latex-QD-antibody complex was collected, washed with storage buffer (2 mM borax, 0.1% BSA, pH 9.0), resuspended in 200 µL storage buffer, and stored in the dark at 4 °C.
TextSentencer_T256 40608-40827 Sentence denotes After centrifugation at 27,237 × g for 5 min, the latex-QD-antibody complex was collected, washed with storage buffer (2 mM borax, 0.1% BSA, pH 9.0), resuspended in 200 µL storage buffer, and stored in the dark at 4 °C.
TextSentencer_T257 40828-40934 Sentence denotes To conjugate mouse serum IgG with latex-QDs, the same molar ratio of mouse serum IgG to antibody was used.
TextSentencer_T257 40828-40934 Sentence denotes To conjugate mouse serum IgG with latex-QDs, the same molar ratio of mouse serum IgG to antibody was used.
TextSentencer_T258 40935-41033 Sentence denotes Nitrocellulose membrane (#HF180, EMD Millipore, Darmstadt, Germany) was used for antibody coating.
TextSentencer_T258 40935-41033 Sentence denotes Nitrocellulose membrane (#HF180, EMD Millipore, Darmstadt, Germany) was used for antibody coating.
TextSentencer_T259 41034-41216 Sentence denotes Conjugate pads (#G028, EMD Millipore), sample pad (#C083, EMD Millipore), and an absorbent pad (#G048, EMD Millipore) were prepared on the backing card (#754104, Rockville, MD, USA).
TextSentencer_T259 41034-41216 Sentence denotes Conjugate pads (#G028, EMD Millipore), sample pad (#C083, EMD Millipore), and an absorbent pad (#G048, EMD Millipore) were prepared on the backing card (#754104, Rockville, MD, USA).
TextSentencer_T260 41217-41334 Sentence denotes The nitrocellulose membrane was coated with 0.2 μg of goat anti-mouse IgG (Life Technology) on the control line (CL).
TextSentencer_T260 41217-41334 Sentence denotes The nitrocellulose membrane was coated with 0.2 μg of goat anti-mouse IgG (Life Technology) on the control line (CL).
TextSentencer_T261 41335-41498 Sentence denotes Two test lines (0.8 μg/lane) were coated with anti-influenza H5 subtype (1C5) (Test line 1: TL 1) and anti-influenza A NP (7304) (Test line 2: TL 2), respectively.
TextSentencer_T261 41335-41498 Sentence denotes Two test lines (0.8 μg/lane) were coated with anti-influenza H5 subtype (1C5) (Test line 1: TL 1) and anti-influenza A NP (7304) (Test line 2: TL 2), respectively.
TextSentencer_T262 41499-41595 Sentence denotes After drying the membrane at 30 °C for two hours, the diagnostic strip was tested by FICT assay.
TextSentencer_T262 41499-41595 Sentence denotes After drying the membrane at 30 °C for two hours, the diagnostic strip was tested by FICT assay.
TextSentencer_T263 41596-41743 Sentence denotes Test strips containing four components each (a sample application pad, a conjugate pad, a nitrocellulose membrane, and an absorbent pad) were used.
TextSentencer_T263 41596-41743 Sentence denotes Test strips containing four components each (a sample application pad, a conjugate pad, a nitrocellulose membrane, and an absorbent pad) were used.
TextSentencer_T264 41744-41805 Sentence denotes The control line (CL) was coated with 2 mg/mL anti-mouse IgG.
TextSentencer_T264 41744-41805 Sentence denotes The control line (CL) was coated with 2 mg/mL anti-mouse IgG.
TextSentencer_T265 41806-41919 Sentence denotes Test line 1 (TL1) was coated with 2.5 mg/mL anti-influenza H5 subtype-specific HA1 mouse monoclonal antibody 1C5.
TextSentencer_T265 41806-41919 Sentence denotes Test line 1 (TL1) was coated with 2.5 mg/mL anti-influenza H5 subtype-specific HA1 mouse monoclonal antibody 1C5.
TextSentencer_T266 41920-42005 Sentence denotes Test line 2 (TL2) was coated with anti-influenza A NP mouse monoclonal antibody 7304.
TextSentencer_T266 41920-42005 Sentence denotes Test line 2 (TL2) was coated with anti-influenza A NP mouse monoclonal antibody 7304.
TextSentencer_T267 42006-42160 Sentence denotes The conjugate pad or each strip was prepared by dropping 2 µL of diluents of two bioconjugates and the strip was preserved in room temperature before use.
TextSentencer_T267 42006-42160 Sentence denotes The conjugate pad or each strip was prepared by dropping 2 µL of diluents of two bioconjugates and the strip was preserved in room temperature before use.
TextSentencer_T268 42161-42274 Sentence denotes For measurements, 75 µL samples and 75 µL lysis buffer were mixed and applied to the sample pad for lateral flow.
TextSentencer_T268 42161-42274 Sentence denotes For measurements, 75 µL samples and 75 µL lysis buffer were mixed and applied to the sample pad for lateral flow.
TextSentencer_T269 42275-42378 Sentence denotes After 20 min, the result was read by the smartphone-based rapid fluorescent diagnostic system (SRDFDS).
TextSentencer_T269 42275-42378 Sentence denotes After 20 min, the result was read by the smartphone-based rapid fluorescent diagnostic system (SRDFDS).
TextSentencer_T270 42379-42482 Sentence denotes During lateral flow on the strip, the influenza virus in the sample reacted with the latex-QD-antibody.
TextSentencer_T270 42379-42482 Sentence denotes During lateral flow on the strip, the influenza virus in the sample reacted with the latex-QD-antibody.
TextSentencer_T271 42483-42636 Sentence denotes The ratio of TL/CL was measured by the smartphone's camera with two fluorescent filters (one filter each for the 580 nm and 650 nm emission wavelengths).
TextSentencer_T271 42483-42636 Sentence denotes The ratio of TL/CL was measured by the smartphone's camera with two fluorescent filters (one filter each for the 580 nm and 650 nm emission wavelengths).
TextSentencer_T272 42637-42881 Sentence denotes A lightweight and compact (18 × 12 × 14 cm 3 ) module for portable fluorescence detection was designed to fit a smartphone (Galaxy S3, Samsung Electronics, Korea) and was fabricated by a 3D printer (Cubicon Single, 3D Cubicon, Seongnam, Korea).
TextSentencer_T272 42637-42881 Sentence denotes A lightweight and compact (18 × 12 × 14 cm 3 ) module for portable fluorescence detection was designed to fit a smartphone (Galaxy S3, Samsung Electronics, Korea) and was fabricated by a 3D printer (Cubicon Single, 3D Cubicon, Seongnam, Korea).
TextSentencer_T273 42882-42978 Sentence denotes A black ABS filament was used to make a light and robust module while reducing light scattering.
TextSentencer_T273 42882-42978 Sentence denotes A black ABS filament was used to make a light and robust module while reducing light scattering.
TextSentencer_T274 42979-43169 Sentence denotes The module included a custom-made parabolic metal-coated light concentrator placed between the specimen strip and smartphone camera for efficient collection of the fluorescence signal [13] .
TextSentencer_T274 42979-43169 Sentence denotes The module included a custom-made parabolic metal-coated light concentrator placed between the specimen strip and smartphone camera for efficient collection of the fluorescence signal [13] .
TextSentencer_T275 43170-43297 Sentence denotes An LED (M365D2, Thorlabs, Newton, NJ, USA) connected to a power supply (LEDD1B, Thorlabs) was used for fluorescence excitation.
TextSentencer_T275 43170-43297 Sentence denotes An LED (M365D2, Thorlabs, Newton, NJ, USA) connected to a power supply (LEDD1B, Thorlabs) was used for fluorescence excitation.
TextSentencer_T276 43298-43527 Sentence denotes An OD4 bandpass filter with a center wavelength of 365 nm (#65-069, Edmund Optics) and OD4 400 nm short pass filter (#84-689, Edmund Optics, Barrington, NJ, USA) were used to cut-off longer wavelengths for crosstalk minimization.
TextSentencer_T276 43298-43527 Sentence denotes An OD4 bandpass filter with a center wavelength of 365 nm (#65-069, Edmund Optics) and OD4 400 nm short pass filter (#84-689, Edmund Optics, Barrington, NJ, USA) were used to cut-off longer wavelengths for crosstalk minimization.
TextSentencer_T277 43528-43720 Sentence denotes Two bandpass emission filters with center wavelengths of 585 nm and 650 nm (ET585/65m, ET650/45x, Chroma, Bellows Falls, VT, USA) were used for separate collection of the fluorescence signals.
TextSentencer_T277 43528-43720 Sentence denotes Two bandpass emission filters with center wavelengths of 585 nm and 650 nm (ET585/65m, ET650/45x, Chroma, Bellows Falls, VT, USA) were used for separate collection of the fluorescence signals.
TextSentencer_T278 43721-43883 Sentence denotes A translational stage with a side-mounted micrometer (XR50P, Thorlabs) was placed on the bottom plate of the module for accurate position control of the specimen.
TextSentencer_T278 43721-43883 Sentence denotes A translational stage with a side-mounted micrometer (XR50P, Thorlabs) was placed on the bottom plate of the module for accurate position control of the specimen.
TextSentencer_T279 43884-43972 Sentence denotes A custom-made plate was fixed on top of the micrometer stage to hold the specimen strip.
TextSentencer_T279 43884-43972 Sentence denotes A custom-made plate was fixed on top of the micrometer stage to hold the specimen strip.
TextSentencer_T280 43973-44143 Sentence denotes The module was designed to ensure convenient switching of the emission optical filters, which were placed between the parabolic metal structure and the smartphone camera.
TextSentencer_T280 43973-44143 Sentence denotes The module was designed to ensure convenient switching of the emission optical filters, which were placed between the parabolic metal structure and the smartphone camera.
TextSentencer_T281 44144-44233 Sentence denotes Once the conjugates were dropped onto the specimen strip, it was inserted onto the stage.
TextSentencer_T281 44144-44233 Sentence denotes Once the conjugates were dropped onto the specimen strip, it was inserted onto the stage.
TextSentencer_T282 44234-44322 Sentence denotes Initial detection was tested on the CL position, which showed fluorescence in all cases.
TextSentencer_T282 44234-44322 Sentence denotes Initial detection was tested on the CL position, which showed fluorescence in all cases.
TextSentencer_T283 44323-44493 Sentence denotes The most accurate position was found by adjusting the actuator of the stage, and the signal data was analyzed using a smartphone application and sent to an online server.
TextSentencer_T283 44323-44493 Sentence denotes The most accurate position was found by adjusting the actuator of the stage, and the signal data was analyzed using a smartphone application and sent to an online server.
TextSentencer_T284 44494-44576 Sentence denotes Analysis of the signal was based on the pictures taken by the smartphone's camera.
TextSentencer_T284 44494-44576 Sentence denotes Analysis of the signal was based on the pictures taken by the smartphone's camera.
TextSentencer_T285 44577-44693 Sentence denotes A mobile app for the smartphone determines the intensity of the whole fluorescent area in addition to the band area.
TextSentencer_T285 44577-44693 Sentence denotes A mobile app for the smartphone determines the intensity of the whole fluorescent area in addition to the band area.
TextSentencer_T286 44694-44840 Sentence denotes The smartphone's camera acquires the collected fluorescent light, and the total intensities are estimated by averaging the pixel intensity values.
TextSentencer_T286 44694-44840 Sentence denotes The smartphone's camera acquires the collected fluorescent light, and the total intensities are estimated by averaging the pixel intensity values.
TextSentencer_T287 44841-45003 Sentence denotes The reflective light concentrator structure with an LED module and the smartphone's camera aperture were placed close to each other as previously described [13] .
TextSentencer_T287 44841-45003 Sentence denotes The reflective light concentrator structure with an LED module and the smartphone's camera aperture were placed close to each other as previously described [13] .
TextSentencer_T288 45004-45077 Sentence denotes The reflectors were coated with aluminum to improve the light reflection.
TextSentencer_T288 45004-45077 Sentence denotes The reflectors were coated with aluminum to improve the light reflection.
TextSentencer_T289 45078-45161 Sentence denotes The reflectivity of aluminum coating was about 90% in the visible wavelength range.
TextSentencer_T289 45078-45161 Sentence denotes The reflectivity of aluminum coating was about 90% in the visible wavelength range.
TextSentencer_T290 45162-45226 Sentence denotes The smartphone used in this study had an 8-million-pixel camera.
TextSentencer_T290 45162-45226 Sentence denotes The smartphone used in this study had an 8-million-pixel camera.
TextSentencer_T291 45227-45418 Sentence denotes Diagnostic results, including positive and negative virus infection decisions based on the cut-off value along with the measurement locations, were automatically posted on an online database.
TextSentencer_T291 45227-45418 Sentence denotes Diagnostic results, including positive and negative virus infection decisions based on the cut-off value along with the measurement locations, were automatically posted on an online database.
TextSentencer_T292 45419-45641 Sentence denotes The proposed diagnostic system was evaluated at the National Institute of Hygiene and Epidemiology (NIHE), Hanoi, Vietnam, with throat swab samples from H5N1-confirmed patients (n = 14) listed by the WHO between 2003-2010.
TextSentencer_T292 45419-45641 Sentence denotes The proposed diagnostic system was evaluated at the National Institute of Hygiene and Epidemiology (NIHE), Hanoi, Vietnam, with throat swab samples from H5N1-confirmed patients (n = 14) listed by the WHO between 2003-2010.
TextSentencer_T293 45642-46099 Sentence denotes For the H5N1-negative control group, Vietnamese patients with an unknown disease (n = 3), respiratory syncytial virus (RSV) (n = 17), metapneumovirus (MPV) (n = 4), bocavirus (HBO) (n = 2), parainfluenza virus (PIV) (n = 5), human rhinovirus (HRV) (n = 5), co-infection of RSV and coronavirus (CoV) (n = 1), CoV NL63 (n = 2), co-infection of human enterovirus (HEV) and CoV (n =1), and a Korean patient with unknown disease (n = 1) were tested using SRDFDS.
TextSentencer_T293 45642-46099 Sentence denotes For the H5N1-negative control group, Vietnamese patients with an unknown disease (n = 3), respiratory syncytial virus (RSV) (n = 17), metapneumovirus (MPV) (n = 4), bocavirus (HBO) (n = 2), parainfluenza virus (PIV) (n = 5), human rhinovirus (HRV) (n = 5), co-infection of RSV and coronavirus (CoV) (n = 1), CoV NL63 (n = 2), co-infection of human enterovirus (HEV) and CoV (n =1), and a Korean patient with unknown disease (n = 1) were tested using SRDFDS.
TextSentencer_T294 46100-46228 Sentence denotes This research was approved by the Wonkwang University Hospital Institutional Review Board (approval no.: WKIRB-201607-HRBR-078).
TextSentencer_T294 46100-46228 Sentence denotes This research was approved by the Wonkwang University Hospital Institutional Review Board (approval no.: WKIRB-201607-HRBR-078).
TextSentencer_T295 46229-46378 Sentence denotes All patients had an acute respiratory infection and agreed to participate in the study, and informed consents were processed before taking specimens.
TextSentencer_T295 46229-46378 Sentence denotes All patients had an acute respiratory infection and agreed to participate in the study, and informed consents were processed before taking specimens.
TextSentencer_T296 46379-46572 Sentence denotes Nucleic acid was extracted from specimens from all patients using the RNeasy Mini kit (Qiagen, Mississauga, ON, Canada) and amplified by commercial multiplex assay (Seegene Inc., Seoul, Korea).
TextSentencer_T296 46379-46572 Sentence denotes Nucleic acid was extracted from specimens from all patients using the RNeasy Mini kit (Qiagen, Mississauga, ON, Canada) and amplified by commercial multiplex assay (Seegene Inc., Seoul, Korea).
TextSentencer_T297 46573-46678 Sentence denotes The R-Mix rapid cell culture method (Quidel, San Diego, CA, USA) was used for the diagnoses of specimens.
TextSentencer_T297 46573-46678 Sentence denotes The R-Mix rapid cell culture method (Quidel, San Diego, CA, USA) was used for the diagnoses of specimens.
TextSentencer_T298 46679-46792 Sentence denotes For the rapid test, patient throat swab samples were directly applied to assays, and results were read in 20 min.
TextSentencer_T298 46679-46792 Sentence denotes For the rapid test, patient throat swab samples were directly applied to assays, and results were read in 20 min.
TextSentencer_T299 46793-46921 Sentence denotes The mean, SD, linear regression, Student's t-test, and one-way analysis of variance (ANOVA) were conducted using GraphPad Prism.
TextSentencer_T299 46793-46921 Sentence denotes The mean, SD, linear regression, Student's t-test, and one-way analysis of variance (ANOVA) were conducted using GraphPad Prism.
TextSentencer_T300 46922-46962 Sentence denotes Results were presented as the mean ± SD.
TextSentencer_T300 46922-46962 Sentence denotes Results were presented as the mean ± SD.
TextSentencer_T301 46963-47044 Sentence denotes Statistical analysis was performed using a one-way ANOVA between multiple groups.
TextSentencer_T301 46963-47044 Sentence denotes Statistical analysis was performed using a one-way ANOVA between multiple groups.
TextSentencer_T302 47045-47092 Sentence denotes A value of P < 0.05 was considered significant.
TextSentencer_T302 47045-47092 Sentence denotes A value of P < 0.05 was considered significant.
TextSentencer_T303 47093-47175 Sentence denotes ROC curves and 95% CIs were computed to confirm assay sensitivity and specificity.
TextSentencer_T303 47093-47175 Sentence denotes ROC curves and 95% CIs were computed to confirm assay sensitivity and specificity.
TextSentencer_T304 47176-47233 Sentence denotes Supplementary figures. http://www.thno.org/v08p6132s1.pdf
TextSentencer_T304 47176-47233 Sentence denotes Supplementary figures. http://www.thno.org/v08p6132s1.pdf