CORD-19-Sentences  

CORD-19:ded7d3ae6f8bbe7866a224433f9aac0471e758ae JSONTXT 9 Projects

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Id Subject Object Predicate Lexical cue
TextSentencer_T1 0-123 Sentence denotes Diagnosis of human respiratory syncytial virus infection using reverse transcription loop-mediated isothermal amplification
TextSentencer_T1 0-123 Sentence denotes Diagnosis of human respiratory syncytial virus infection using reverse transcription loop-mediated isothermal amplification
TextSentencer_T2 125-133 Sentence denotes Abstract
TextSentencer_T2 125-133 Sentence denotes Abstract
TextSentencer_T3 134-267 Sentence denotes Human respiratory syncytial virus (RSV) is a major causative agent of lower respiratory tract infections in children and the elderly.
TextSentencer_T3 134-267 Sentence denotes Human respiratory syncytial virus (RSV) is a major causative agent of lower respiratory tract infections in children and the elderly.
TextSentencer_T4 268-474 Sentence denotes A reverse transcription-loop-mediated isothermal amplification (RT-LAMP) was developed assay to amplify the genome of RSV subgroups A and B, in order to improve current diagnostic methods for RSV infection.
TextSentencer_T4 268-474 Sentence denotes A reverse transcription-loop-mediated isothermal amplification (RT-LAMP) was developed assay to amplify the genome of RSV subgroups A and B, in order to improve current diagnostic methods for RSV infection.
TextSentencer_T5 475-648 Sentence denotes The primer sets for RT-LAMP were designed using highly conserved nucleotide sequences in the matrix protein region of subgroups A and B, and were specific for each subgroup.
TextSentencer_T5 475-648 Sentence denotes The primer sets for RT-LAMP were designed using highly conserved nucleotide sequences in the matrix protein region of subgroups A and B, and were specific for each subgroup.
TextSentencer_T6 649-891 Sentence denotes The RT-LAMP efficiency was compared to virus isolation and a commercially available enzyme immunoassay (EIA) for RSV detection (BD Directigen EZ RSV test TM ), using nasopharyngeal aspirates from 59 children with respiratory tract infections.
TextSentencer_T6 649-891 Sentence denotes The RT-LAMP efficiency was compared to virus isolation and a commercially available enzyme immunoassay (EIA) for RSV detection (BD Directigen EZ RSV test TM ), using nasopharyngeal aspirates from 59 children with respiratory tract infections.
TextSentencer_T7 892-974 Sentence denotes The RT-LAMP was specific for RSV and could not detect other respiratory pathogens.
TextSentencer_T7 892-974 Sentence denotes The RT-LAMP was specific for RSV and could not detect other respiratory pathogens.
TextSentencer_T8 975-1083 Sentence denotes 61% (36/59) of children were positive by RT-LAMP, 34% (20/59) by viral isolation, and 56% (26/46) by EZ RSV.
TextSentencer_T8 975-1083 Sentence denotes 61% (36/59) of children were positive by RT-LAMP, 34% (20/59) by viral isolation, and 56% (26/46) by EZ RSV.
TextSentencer_T9 1084-1201 Sentence denotes Of 16 specimens that were negative by both antigen detection and virus isolation, 12.5% (2/16) were RT-LAMP positive.
TextSentencer_T9 1084-1201 Sentence denotes Of 16 specimens that were negative by both antigen detection and virus isolation, 12.5% (2/16) were RT-LAMP positive.
TextSentencer_T10 1202-1297 Sentence denotes These results suggest that the RT-LAMP is more sensitive than other methods used to detect RSV.
TextSentencer_T10 1202-1297 Sentence denotes These results suggest that the RT-LAMP is more sensitive than other methods used to detect RSV.
TextSentencer_T11 1298-1414 Sentence denotes The RT-LAMP assay developed in this study may be useful for diagnostic and epidemiological studies of RSV infection.
TextSentencer_T11 1298-1414 Sentence denotes The RT-LAMP assay developed in this study may be useful for diagnostic and epidemiological studies of RSV infection.
TextSentencer_T12 1416-1636 Sentence denotes Human respiratory syncytial virus (RSV) belongs to the genus Pneumovirus in the subfamily Pneumovirinae of the family Paramyxoviridae, and is a major causative agent of respiratory tract infections in children worldwide.
TextSentencer_T12 1416-1636 Sentence denotes Human respiratory syncytial virus (RSV) belongs to the genus Pneumovirus in the subfamily Pneumovirinae of the family Paramyxoviridae, and is a major causative agent of respiratory tract infections in children worldwide.
TextSentencer_T13 1637-1898 Sentence denotes Children born preterm or with underlying cardiopulmonary disorders are at especially high risk of developing severe and lethal RSV respiratory tract infections (American Academy of Pediatrics, 2003; Girard et al., 2005; Saijo et al., 1993 Saijo et al., , 1994 .
TextSentencer_T13 1637-1898 Sentence denotes Children born preterm or with underlying cardiopulmonary disorders are at especially high risk of developing severe and lethal RSV respiratory tract infections (American Academy of Pediatrics, 2003; Girard et al., 2005; Saijo et al., 1993 Saijo et al., , 1994 .
TextSentencer_T14 1899-2169 Sentence denotes Elderly and immunocompromised individuals are also susceptible to severe respiratory diseases, highlighting the importance of preventing nasocomial and in-house RSV infections in hospitals and care centers for the elderly (Englund et al., 1988; Falsey and Walsh, 2000) .
TextSentencer_T14 1899-2169 Sentence denotes Elderly and immunocompromised individuals are also susceptible to severe respiratory diseases, highlighting the importance of preventing nasocomial and in-house RSV infections in hospitals and care centers for the elderly (Englund et al., 1988; Falsey and Walsh, 2000) .
TextSentencer_T15 2170-2384 Sentence denotes Palivizumab, a monoclonal antibody with neutralizing activity against RSV, was developed recently and made available commercially as an effective prophylaxis against disease (American Academy of Pediatrics, 2003) .
TextSentencer_T15 2170-2384 Sentence denotes Palivizumab, a monoclonal antibody with neutralizing activity against RSV, was developed recently and made available commercially as an effective prophylaxis against disease (American Academy of Pediatrics, 2003) .
TextSentencer_T16 2385-2487 Sentence denotes Thus, precise diagnostic and epidemiological tests are important for the prevention of RSV infections.
TextSentencer_T16 2385-2487 Sentence denotes Thus, precise diagnostic and epidemiological tests are important for the prevention of RSV infections.
TextSentencer_T17 2488-2684 Sentence denotes The diagnosis of RSV infection is generally undertaken by virus isolation and/or antigen detection using an enzyme immunoassay (EIA) (Dayan et al., 2002; Falsey et al., 2002; Reina et al., 2004) .
TextSentencer_T17 2488-2684 Sentence denotes The diagnosis of RSV infection is generally undertaken by virus isolation and/or antigen detection using an enzyme immunoassay (EIA) (Dayan et al., 2002; Falsey et al., 2002; Reina et al., 2004) .
TextSentencer_T18 2685-2865 Sentence denotes However, because of the thermolability of RSV, the rate of isolation is dependent on conditions such as sample collection and cold chain from sample collection to cell inoculation.
TextSentencer_T18 2685-2865 Sentence denotes However, because of the thermolability of RSV, the rate of isolation is dependent on conditions such as sample collection and cold chain from sample collection to cell inoculation.
TextSentencer_T19 2866-3142 Sentence denotes EIAs are useful as bed-side diagnostic tools in clinical settings, however their sensitivity and specificity are generally between 70 and 90%, suggesting that misdiagnosis of RSV infection is common (10-30%) (Grondahl et al., 2005; Ohm-Smith et al., 2004; Wren et al., 1990) .
TextSentencer_T19 2866-3142 Sentence denotes EIAs are useful as bed-side diagnostic tools in clinical settings, however their sensitivity and specificity are generally between 70 and 90%, suggesting that misdiagnosis of RSV infection is common (10-30%) (Grondahl et al., 2005; Ohm-Smith et al., 2004; Wren et al., 1990) .
TextSentencer_T20 3143-3374 Sentence denotes In addition, it is difficult to detect RSV antigen in nasopharyngeal aspirates collected from adults, since only a small amount of RSV antigen can be found (Casiano-Colon et al., 2003; Falsey et al., 2002; Falsey and Walsh, 2000) .
TextSentencer_T20 3143-3374 Sentence denotes In addition, it is difficult to detect RSV antigen in nasopharyngeal aspirates collected from adults, since only a small amount of RSV antigen can be found (Casiano-Colon et al., 2003; Falsey et al., 2002; Falsey and Walsh, 2000) .
TextSentencer_T21 3375-3497 Sentence denotes The development of more sensitive and specific detection methods is therefore essential for more precise diagnoses of RSV.
TextSentencer_T21 3375-3497 Sentence denotes The development of more sensitive and specific detection methods is therefore essential for more precise diagnoses of RSV.
TextSentencer_T22 3498-3719 Sentence denotes As a result, molecular methods such as reverse transcription (RT)-polymerase chain reaction (PCR) and quantitative real-time RT-PCR have been developed for the detection of RSV (Falsey et al., 2002 Gilbert et al., 1996) .
TextSentencer_T22 3498-3719 Sentence denotes As a result, molecular methods such as reverse transcription (RT)-polymerase chain reaction (PCR) and quantitative real-time RT-PCR have been developed for the detection of RSV (Falsey et al., 2002 Gilbert et al., 1996) .
TextSentencer_T23 3720-3908 Sentence denotes Loop-mediated isothermal amplification (LAMP) was developed recently to amplify specific nucleic acid sequences using a designated primer set (Nagamine et al., 2002; Notomi et al., 2000) .
TextSentencer_T23 3720-3908 Sentence denotes Loop-mediated isothermal amplification (LAMP) was developed recently to amplify specific nucleic acid sequences using a designated primer set (Nagamine et al., 2002; Notomi et al., 2000) .
TextSentencer_T24 3909-4202 Sentence denotes Amplification by LAMP is simple, since the reaction mixture is incubated at a single temperature for less than 1 h, and specificity is high because at least six specific primer region sequences are required for construction of the LAMP primer set (Nagamine et al., 2002; Notomi et al., 2000) .
TextSentencer_T24 3909-4202 Sentence denotes Amplification by LAMP is simple, since the reaction mixture is incubated at a single temperature for less than 1 h, and specificity is high because at least six specific primer region sequences are required for construction of the LAMP primer set (Nagamine et al., 2002; Notomi et al., 2000) .
TextSentencer_T25 4203-4416 Sentence denotes The amplified product level can be monitored in a real-time quantitative manner by measuring turbidity since nucleic acid amplification results in the precipitation of magnesium pyrophosphate (Mori et al., 2001) .
TextSentencer_T25 4203-4416 Sentence denotes The amplified product level can be monitored in a real-time quantitative manner by measuring turbidity since nucleic acid amplification results in the precipitation of magnesium pyrophosphate (Mori et al., 2001) .
TextSentencer_T26 4417-4556 Sentence denotes The LAMP assay can also be used with RNA templates by combining reverse transcription reactions with LAMP (RT-LAMP) (Notomi et al., 2000) .
TextSentencer_T26 4417-4556 Sentence denotes The LAMP assay can also be used with RNA templates by combining reverse transcription reactions with LAMP (RT-LAMP) (Notomi et al., 2000) .
TextSentencer_T27 4557-4797 Sentence denotes Genetic diagnosis by RT-LAMP has been developed for infections caused by RNA viruses, such as severe acute respiratory syndrome (SARS), West Nile encephalitis, and Dengue fever (Hong et al., 2004; Parida et al., 2004 Parida et al., , 2005 .
TextSentencer_T27 4557-4797 Sentence denotes Genetic diagnosis by RT-LAMP has been developed for infections caused by RNA viruses, such as severe acute respiratory syndrome (SARS), West Nile encephalitis, and Dengue fever (Hong et al., 2004; Parida et al., 2004 Parida et al., , 2005 .
TextSentencer_T28 4798-4993 Sentence denotes Ushio et al. (2005) recently developed RT-LAMP for detection of the RSV genome and the primers used in their method were designed according to the nucleotide sequence of the nucleoprotein region.
TextSentencer_T28 4798-4993 Sentence denotes Ushio et al. (2005) recently developed RT-LAMP for detection of the RSV genome and the primers used in their method were designed according to the nucleotide sequence of the nucleoprotein region.
TextSentencer_T29 4994-5183 Sentence denotes Highly conserved sequences of the nucleotide sequence are located in the matrix protein region of RSV isolates, suggesting that this region was a suitable target for designing LAMP primers.
TextSentencer_T29 4994-5183 Sentence denotes Highly conserved sequences of the nucleotide sequence are located in the matrix protein region of RSV isolates, suggesting that this region was a suitable target for designing LAMP primers.
TextSentencer_T30 5184-5380 Sentence denotes In the present study, a RT-LAMP method was developed in which the LAMP primers were designed from the matrix protein gene, and the competence of this assay was compared to virus isolation and EIA.
TextSentencer_T30 5184-5380 Sentence denotes In the present study, a RT-LAMP method was developed in which the LAMP primers were designed from the matrix protein gene, and the competence of this assay was compared to virus isolation and EIA.
TextSentencer_T31 5381-5548 Sentence denotes 59 nasopharyngeal aspirates were collected from children with respiratory tract infections from November 2004 to November 2005 at Sendai Medical Center, Sendai, Japan.
TextSentencer_T31 5381-5548 Sentence denotes 59 nasopharyngeal aspirates were collected from children with respiratory tract infections from November 2004 to November 2005 at Sendai Medical Center, Sendai, Japan.
TextSentencer_T32 5549-5806 Sentence denotes Nasopharyngeal aspirates were suspended in transport medium containing Eagle's minimum essential medium (MEM, Invitrogen Corp., Carlsbad, CA, USA) supplemented with 0.5% of gelatin, and antibiotics (Numazaki et al., 1987) and stored at −20 • C prior to use.
TextSentencer_T32 5549-5806 Sentence denotes Nasopharyngeal aspirates were suspended in transport medium containing Eagle's minimum essential medium (MEM, Invitrogen Corp., Carlsbad, CA, USA) supplemented with 0.5% of gelatin, and antibiotics (Numazaki et al., 1987) and stored at −20 • C prior to use.
TextSentencer_T33 5807-5926 Sentence denotes The nasopharyngeal aspirates were centrifuged at 3000 rpm for 15 min, and the supernatant was used for virus isolation.
TextSentencer_T33 5807-5926 Sentence denotes The nasopharyngeal aspirates were centrifuged at 3000 rpm for 15 min, and the supernatant was used for virus isolation.
TextSentencer_T34 5927-6128 Sentence denotes Detection of RSV antigen was analyzed in 46 of the 59 nasopharyngeal aspirates using the enzyme immunoassay (EIA), BD Directigen EZ RSV test TM (Becton, Dickinson and Company, Franklin Lakes, NJ, USA).
TextSentencer_T34 5927-6128 Sentence denotes Detection of RSV antigen was analyzed in 46 of the 59 nasopharyngeal aspirates using the enzyme immunoassay (EIA), BD Directigen EZ RSV test TM (Becton, Dickinson and Company, Franklin Lakes, NJ, USA).
TextSentencer_T35 6129-6192 Sentence denotes The nasopharyngeal aspirates were also used for RNA extraction.
TextSentencer_T35 6129-6192 Sentence denotes The nasopharyngeal aspirates were also used for RNA extraction.
TextSentencer_T36 6193-6418 Sentence denotes Virus isolation was performed using six kinds of cells, MDCK, HEp-2, Vero, HMV-II, LLC-MK2, and human embryo fibroblasts, by seeding them in 96-well plate as described previously (Numazaki et al., 1987; Suzuki et al., 2005) .
TextSentencer_T36 6193-6418 Sentence denotes Virus isolation was performed using six kinds of cells, MDCK, HEp-2, Vero, HMV-II, LLC-MK2, and human embryo fibroblasts, by seeding them in 96-well plate as described previously (Numazaki et al., 1987; Suzuki et al., 2005) .
TextSentencer_T37 6419-6585 Sentence denotes The RSV strains used in this study were laboratory strains (Long and CH-18537) and wild type viruses isolated in 1996 (N96-77, N97-23, N91-730, N96-51, and N96-728) .
TextSentencer_T37 6419-6585 Sentence denotes The RSV strains used in this study were laboratory strains (Long and CH-18537) and wild type viruses isolated in 1996 (N96-77, N97-23, N91-730, N96-51, and N96-728) .
TextSentencer_T38 6586-6934 Sentence denotes Human metapneumovirus was isolated in the Virus Research Center at Sendai Medical Center (hMPV; 2003-2404), human coronaviruses (HCoV) strain NL63 was supplied by Dr. Hoek (University of Amsterdam, the Netherlands), HCoV strain 229E was supplied by Dr. Nomura (FUJITA Health University, Japan), SARS coronavirus strain Frankfurt was supplied by Dr.
TextSentencer_T38 6586-6934 Sentence denotes Human metapneumovirus was isolated in the Virus Research Center at Sendai Medical Center (hMPV; 2003-2404), human coronaviruses (HCoV) strain NL63 was supplied by Dr. Hoek (University of Amsterdam, the Netherlands), HCoV strain 229E was supplied by Dr. Nomura (FUJITA Health University, Japan), SARS coronavirus strain Frankfurt was supplied by Dr.
TextSentencer_T39 6935-6937 Sentence denotes J.
TextSentencer_T39 6935-6937 Sentence denotes J.
TextSentencer_T40 6938-7366 Sentence denotes Ziebuhr (University of Würzburg, Germany), parainfluenza virus 3 was purchased from the American Tissue Culture Collection (ATCC) (C-245), influenza viruses strains A:H5N1, A:H3N2, A:H1N1, and B were stored in the National Institute of Infectious Diseases in Japan, and adenoviruses serotype 3, strain O.B., serotype 4, strain R163, and serotype 7, strain Gomen were stored in National Institute of Infectious Diseases in Japan.
TextSentencer_T40 6938-7366 Sentence denotes Ziebuhr (University of Würzburg, Germany), parainfluenza virus 3 was purchased from the American Tissue Culture Collection (ATCC) (C-245), influenza viruses strains A:H5N1, A:H3N2, A:H1N1, and B were stored in the National Institute of Infectious Diseases in Japan, and adenoviruses serotype 3, strain O.B., serotype 4, strain R163, and serotype 7, strain Gomen were stored in National Institute of Infectious Diseases in Japan.
TextSentencer_T41 7367-7629 Sentence denotes Bacterial pathogens that induce respiratory tract infections, like Bordetella pertussis (Tohama), Bordetella parapertussis (ATCC15311), Haemophils influenza biotype 3, Haemophils parainfluenza biotype 1, and Mycoplasma pneumonia (M129), were also stored in NIID.
TextSentencer_T41 7367-7629 Sentence denotes Bacterial pathogens that induce respiratory tract infections, like Bordetella pertussis (Tohama), Bordetella parapertussis (ATCC15311), Haemophils influenza biotype 3, Haemophils parainfluenza biotype 1, and Mycoplasma pneumonia (M129), were also stored in NIID.
TextSentencer_T42 7630-7759 Sentence denotes RSV was propagated in HEp-2 cells and titrated using the standard plaque assay on HEp-2 cells cultured in 12-well culture plates.
TextSentencer_T42 7630-7759 Sentence denotes RSV was propagated in HEp-2 cells and titrated using the standard plaque assay on HEp-2 cells cultured in 12-well culture plates.
TextSentencer_T43 7760-7887 Sentence denotes RSV strains are divided into two subgroups (A and B) based on antigenic variation (Gimenez et al., 1986; Mufson et al., 1985) .
TextSentencer_T43 7760-7887 Sentence denotes RSV strains are divided into two subgroups (A and B) based on antigenic variation (Gimenez et al., 1986; Mufson et al., 1985) .
TextSentencer_T44 7888-8024 Sentence denotes The primer sets for the RT-LAMP assay were designed from the alignment sequences of three subgroup A RSV strains (GenBank accession Nos.
TextSentencer_T44 7888-8024 Sentence denotes The primer sets for the RT-LAMP assay were designed from the alignment sequences of three subgroup A RSV strains (GenBank accession Nos.
TextSentencer_T45 8025-8220 Sentence denotes NC 001803, U39662, U50362) and three subgroup B RSV strains (NC 001781, AF013254, and AY353550) using the online LAMP primer design software (PrimerExplorer; http://primerexplorer.jp/index.html).
TextSentencer_T45 8025-8220 Sentence denotes NC 001803, U39662, U50362) and three subgroup B RSV strains (NC 001781, AF013254, and AY353550) using the online LAMP primer design software (PrimerExplorer; http://primerexplorer.jp/index.html).
TextSentencer_T46 8221-8278 Sentence denotes The loop primer set was also designed with this software.
TextSentencer_T46 8221-8278 Sentence denotes The loop primer set was also designed with this software.
TextSentencer_T47 8279-8369 Sentence denotes The RSV RT-LAMP primer set described by Ushio et al. (2005) was also used for experiments.
TextSentencer_T47 8279-8369 Sentence denotes The RSV RT-LAMP primer set described by Ushio et al. (2005) was also used for experiments.
TextSentencer_T48 8370-8486 Sentence denotes RNA was extracted from virus stocks using TRIzol reagent (Invitrogen), according to the manufacturer's instructions.
TextSentencer_T48 8370-8486 Sentence denotes RNA was extracted from virus stocks using TRIzol reagent (Invitrogen), according to the manufacturer's instructions.
TextSentencer_T49 8487-8641 Sentence denotes DNA was extracted from stocks of adenovirus or bacteria using QIA-GEN Genomic-tips (Qiagen, Hilden, Germany) according to the manufacturer's instructions.
TextSentencer_T49 8487-8641 Sentence denotes DNA was extracted from stocks of adenovirus or bacteria using QIA-GEN Genomic-tips (Qiagen, Hilden, Germany) according to the manufacturer's instructions.
TextSentencer_T50 8642-8785 Sentence denotes RNA was extracted from 140 l of each nasopharyngeal aspirate and eluted with 50 l of elute buffer using the QIAamp Viral RNA Mini Kit (Qiagen).
TextSentencer_T50 8642-8785 Sentence denotes RNA was extracted from 140 l of each nasopharyngeal aspirate and eluted with 50 l of elute buffer using the QIAamp Viral RNA Mini Kit (Qiagen).
TextSentencer_T51 8786-8855 Sentence denotes Total RNA and genomic DNA were quantitated by measuring the OD value.
TextSentencer_T51 8786-8855 Sentence denotes Total RNA and genomic DNA were quantitated by measuring the OD value.
TextSentencer_T52 8856-8981 Sentence denotes The RT-LAMP assay was performed using the Loopamp RNA amplification reagents kit (RT-LAMP; Eiken, Tokyo, Japan) as described.
TextSentencer_T52 8856-8981 Sentence denotes The RT-LAMP assay was performed using the Loopamp RNA amplification reagents kit (RT-LAMP; Eiken, Tokyo, Japan) as described.
TextSentencer_T53 8982-9259 Sentence denotes 5 l of each sample of extracted RNA or DNA was mixed with 40 pmol of the FIP and BIP primers, 20 pmol of the Loop-F and Loop-B primers, 5 pmol of the F3 and B3 primers, 1 l of Enzyme Mix, and 12.5 l of Reaction Mix, and distilled water was added to make a final volume of 25 l.
TextSentencer_T53 8982-9259 Sentence denotes 5 l of each sample of extracted RNA or DNA was mixed with 40 pmol of the FIP and BIP primers, 20 pmol of the Loop-F and Loop-B primers, 5 pmol of the F3 and B3 primers, 1 l of Enzyme Mix, and 12.5 l of Reaction Mix, and distilled water was added to make a final volume of 25 l.
TextSentencer_T54 9260-9426 Sentence denotes For real-time monitoring of the RT-LAMP assay, the reaction mixture was incubated at 63 • C for 45 min in the Loopamp real-time turbidimeter Teramecs, Kyoto, Japan) .
TextSentencer_T54 9260-9426 Sentence denotes For real-time monitoring of the RT-LAMP assay, the reaction mixture was incubated at 63 • C for 45 min in the Loopamp real-time turbidimeter Teramecs, Kyoto, Japan) .
TextSentencer_T55 9427-9485 Sentence denotes A no template negative control was included in each assay.
TextSentencer_T55 9427-9485 Sentence denotes A no template negative control was included in each assay.
TextSentencer_T56 9486-9633 Sentence denotes For the purposes of this study, samples in which the magnitude of the turbidity signal was greater than 0.1 within 45 min were considered positive.
TextSentencer_T56 9486-9633 Sentence denotes For the purposes of this study, samples in which the magnitude of the turbidity signal was greater than 0.1 within 45 min were considered positive.
TextSentencer_T57 9634-9734 Sentence denotes Time during which the turbidity was greater than 0.1 was expressed as time of positivity in minutes.
TextSentencer_T57 9634-9734 Sentence denotes Time during which the turbidity was greater than 0.1 was expressed as time of positivity in minutes.
TextSentencer_T58 9735-9853 Sentence denotes The minimum copy number of the target regions produced using the RT-LAMP assay was determined from the RT-PCR product.
TextSentencer_T58 9735-9853 Sentence denotes The minimum copy number of the target regions produced using the RT-LAMP assay was determined from the RT-PCR product.
TextSentencer_T59 9854-9986 Sentence denotes Target regions of the RT-LAMP assay were amplified with specific primers and Platinum Taq High Fidelity DNA polymerase (Invitrogen).
TextSentencer_T59 9854-9986 Sentence denotes Target regions of the RT-LAMP assay were amplified with specific primers and Platinum Taq High Fidelity DNA polymerase (Invitrogen).
TextSentencer_T60 9987-10131 Sentence denotes The PCR products were gel-purified with a QIAquick Gel Purification Kit (Qiagen) and the concentration was determined by measuring the OD value.
TextSentencer_T60 9987-10131 Sentence denotes The PCR products were gel-purified with a QIAquick Gel Purification Kit (Qiagen) and the concentration was determined by measuring the OD value.
TextSentencer_T61 10132-10274 Sentence denotes The DNA copy number was calculated using the following formula: copies (number/ml) = ((6.023 × l0 23 × 5 × l0 −5 × OD 260 )/molecular weight).
TextSentencer_T61 10132-10274 Sentence denotes The DNA copy number was calculated using the following formula: copies (number/ml) = ((6.023 × l0 23 × 5 × l0 −5 × OD 260 )/molecular weight).
TextSentencer_T62 10275-10389 Sentence denotes RT-LAMP assay was performed using serially diluted target DNA, and the copy number detection limit was determined.
TextSentencer_T62 10275-10389 Sentence denotes RT-LAMP assay was performed using serially diluted target DNA, and the copy number detection limit was determined.
TextSentencer_T63 10390-10478 Sentence denotes One-step RT-PCR was also carried out to detect RSV from clinical specimens as described.
TextSentencer_T63 10390-10478 Sentence denotes One-step RT-PCR was also carried out to detect RSV from clinical specimens as described.
TextSentencer_T64 10479-10804 Sentence denotes 5 l of each sample was mixed with 1 l of each primer set (sense: GARGTRGCTCCAGAATAYAGGCAT, anti-sense: AGCAT-CACTTGCCCTGAACCATAG; Osiowy, 1998) , 25.0 l of 2× AccessQuik RT-PCR Master Mix (Promega Corporation, Madison, WI, USA), and 1 l of supplemented RT enzyme, and distilled water was added to make a final volume of 50 l.
TextSentencer_T64 10479-10804 Sentence denotes 5 l of each sample was mixed with 1 l of each primer set (sense: GARGTRGCTCCAGAATAYAGGCAT, anti-sense: AGCAT-CACTTGCCCTGAACCATAG; Osiowy, 1998) , 25.0 l of 2× AccessQuik RT-PCR Master Mix (Promega Corporation, Madison, WI, USA), and 1 l of supplemented RT enzyme, and distilled water was added to make a final volume of 50 l.
TextSentencer_T65 10805-11070 Sentence denotes The amplification program consisted of an RT step at 45 • C for 45 min, followed by a denature step at 94 • C for 2 min, and 30 cycles of 93 • C for 30 s, 55 • C for 30 s, and 72 • C for 50 s, with a final extension step of 72 • C for 7 min (Gruteke et al., 2004) .
TextSentencer_T65 10805-11070 Sentence denotes The amplification program consisted of an RT step at 45 • C for 45 min, followed by a denature step at 94 • C for 2 min, and 30 cycles of 93 • C for 30 s, 55 • C for 30 s, and 72 • C for 50 s, with a final extension step of 72 • C for 7 min (Gruteke et al., 2004) .
TextSentencer_T66 11071-11242 Sentence denotes After amplification, amplicons were separated by electrophoresis on a 2% agarose gel, stained with ethidium bromide, and the bands were visualized under ultraviolet light.
TextSentencer_T66 11071-11242 Sentence denotes After amplification, amplicons were separated by electrophoresis on a 2% agarose gel, stained with ethidium bromide, and the bands were visualized under ultraviolet light.
TextSentencer_T67 11243-11475 Sentence denotes RT-LAMP products were purified using the QIAquick PCR Purification Kit (Qiagen), and the purified amplicons were digested with the restriction enzyme, NlaIII or XbaI, under optimal conditions (New England Biolabs, Ipswich, MA, USA).
TextSentencer_T67 11243-11475 Sentence denotes RT-LAMP products were purified using the QIAquick PCR Purification Kit (Qiagen), and the purified amplicons were digested with the restriction enzyme, NlaIII or XbaI, under optimal conditions (New England Biolabs, Ipswich, MA, USA).
TextSentencer_T68 11476-11657 Sentence denotes After 2 h of digestion, RT-LAMP products were separated by electrophoresis on a 3% agarose gel, stained with ethidium bromide, and the bands were visualized under ultraviolet light.
TextSentencer_T68 11476-11657 Sentence denotes After 2 h of digestion, RT-LAMP products were separated by electrophoresis on a 3% agarose gel, stained with ethidium bromide, and the bands were visualized under ultraviolet light.
TextSentencer_T69 11658-11771 Sentence denotes The amplicon from subgroup A RSV was digested with NlaIII, while that from subgroup B RSV was digested with XbaI.
TextSentencer_T69 11658-11771 Sentence denotes The amplicon from subgroup A RSV was digested with NlaIII, while that from subgroup B RSV was digested with XbaI.
TextSentencer_T70 11772-11829 Sentence denotes The primer sets used in this study are shown in Table 1 .
TextSentencer_T70 11772-11829 Sentence denotes The primer sets used in this study are shown in Table 1 .
TextSentencer_T71 11830-11937 Sentence denotes The primer set for RT-LAMP was first designed with the aim of amplifying both the A and B subgroups of RSV.
TextSentencer_T71 11830-11937 Sentence denotes The primer set for RT-LAMP was first designed with the aim of amplifying both the A and B subgroups of RSV.
TextSentencer_T72 11938-12045 Sentence denotes This proved difficult to design, however, because the LAMP assay uses at least six specific primer regions.
TextSentencer_T72 11938-12045 Sentence denotes This proved difficult to design, however, because the LAMP assay uses at least six specific primer regions.
TextSentencer_T73 12046-12166 Sentence denotes Thus, primer sets were designed that had specificity for subgroup A RSV (primer set A) or subgroup B RSV (primer set B).
TextSentencer_T73 12046-12166 Sentence denotes Thus, primer sets were designed that had specificity for subgroup A RSV (primer set A) or subgroup B RSV (primer set B).
TextSentencer_T74 12167-12339 Sentence denotes Based on alignment analysis of RSV genomic sequences, highly conserved sequences in the matrix protein coding region were selected as the target genomic sequence (Fig. 1) .
TextSentencer_T74 12167-12339 Sentence denotes Based on alignment analysis of RSV genomic sequences, highly conserved sequences in the matrix protein coding region were selected as the target genomic sequence (Fig. 1) .
TextSentencer_T75 12340-12520 Sentence denotes A negative control using water was analyzed by RT-LAMP for 60 min to define the value of the turbidity signal that would discriminate positives from negatives in the RT-LAMP assay.
TextSentencer_T75 12340-12520 Sentence denotes A negative control using water was analyzed by RT-LAMP for 60 min to define the value of the turbidity signal that would discriminate positives from negatives in the RT-LAMP assay.
TextSentencer_T76 12521-12630 Sentence denotes None of the negative controls had a turbidity signal that was greater than 0.1 within a 45 min reaction time.
TextSentencer_T76 12521-12630 Sentence denotes None of the negative controls had a turbidity signal that was greater than 0.1 within a 45 min reaction time.
TextSentencer_T77 12631-12727 Sentence denotes Thus, turbid- ity values exceeding 0.1 within the 45 min reaction time were defined as positive.
TextSentencer_T77 12631-12727 Sentence denotes Thus, turbid- ity values exceeding 0.1 within the 45 min reaction time were defined as positive.
TextSentencer_T78 12728-12831 Sentence denotes Specific reactions of the designed primer set were tested on various respiratory pathogens ( Table 2 ).
TextSentencer_T78 12728-12831 Sentence denotes Specific reactions of the designed primer set were tested on various respiratory pathogens ( Table 2 ).
TextSentencer_T79 12832-12923 Sentence denotes The RT-LAMP assay was performed using 100 ng of total RNA or 1 ng of extracted genomic DNA.
TextSentencer_T79 12832-12923 Sentence denotes The RT-LAMP assay was performed using 100 ng of total RNA or 1 ng of extracted genomic DNA.
TextSentencer_T80 12924-13045 Sentence denotes The RT-LAMP using primer set A and primer set B were only able to detect subgroup A RSV and subgroup B RSV, respectively.
TextSentencer_T80 12924-13045 Sentence denotes The RT-LAMP using primer set A and primer set B were only able to detect subgroup A RSV and subgroup B RSV, respectively.
TextSentencer_T81 13046-13230 Sentence denotes Samples prepared from other respiratory pathogens, including human coronaviruses, influenza viruses, metapneumovirus, adenoviruses, and bacteria, showed a negative reaction by RT-LAMP.
TextSentencer_T81 13046-13230 Sentence denotes Samples prepared from other respiratory pathogens, including human coronaviruses, influenza viruses, metapneumovirus, adenoviruses, and bacteria, showed a negative reaction by RT-LAMP.
TextSentencer_T82 13231-13372 Sentence denotes Importantly, these pathogens were positive by PCR or RT-PCR when primer sets were used that were specific for each organism (data not shown).
TextSentencer_T82 13231-13372 Sentence denotes Importantly, these pathogens were positive by PCR or RT-PCR when primer sets were used that were specific for each organism (data not shown).
TextSentencer_T83 13373-13448 Sentence denotes Thus, the primer set described in this report specifically reacts with RSV.
TextSentencer_T83 13373-13448 Sentence denotes Thus, the primer set described in this report specifically reacts with RSV.
TextSentencer_T84 13449-13644 Sentence denotes Amplicons were digested with the restriction enzyme NlaIII or XbaI and separated by electrophoresis on a 3% agarose gel to confirm specific amplification and differentiation of subgroups A and B.
TextSentencer_T84 13449-13644 Sentence denotes Amplicons were digested with the restriction enzyme NlaIII or XbaI and separated by electrophoresis on a 3% agarose gel to confirm specific amplification and differentiation of subgroups A and B.
TextSentencer_T85 13645-13837 Sentence denotes As predicted, the RT-LAMP products amplified from subgroup A RSV were only digested with NlaIII, while the products amplified from subgroup B RSV were only digested with XbaI (data not shown).
TextSentencer_T85 13645-13837 Sentence denotes As predicted, the RT-LAMP products amplified from subgroup A RSV were only digested with NlaIII, while the products amplified from subgroup B RSV were only digested with XbaI (data not shown).
TextSentencer_T86 13838-13957 Sentence denotes The sensitivity of RT-LAMP was next evaluated using serially diluted RNA samples prepared from an RSV stock (Table 3 ).
TextSentencer_T86 13838-13957 Sentence denotes The sensitivity of RT-LAMP was next evaluated using serially diluted RNA samples prepared from an RSV stock (Table 3 ).
TextSentencer_T87 13958-14064 Sentence denotes The infectivity of the RSV stock was determined by plaque assay using HEp-2 cells prior to RNA extraction.
TextSentencer_T87 13958-14064 Sentence denotes The infectivity of the RSV stock was determined by plaque assay using HEp-2 cells prior to RNA extraction.
TextSentencer_T88 14065-14237 Sentence denotes The entire process of preparing the samples including virus propagation, virus titration, RNA extraction, and the RT-LAMP assay, were carried out three times independently.
TextSentencer_T88 14065-14237 Sentence denotes The entire process of preparing the samples including virus propagation, virus titration, RNA extraction, and the RT-LAMP assay, were carried out three times independently.
TextSentencer_T89 14238-14338 Sentence denotes 10 −1 to 10 −2 PFU of RSV was required to provide enough RNA for detection using the RT- LAMP assay.
TextSentencer_T89 14238-14338 Sentence denotes 10 −1 to 10 −2 PFU of RSV was required to provide enough RNA for detection using the RT- LAMP assay.
TextSentencer_T90 14339-14456 Sentence denotes The detection limit of nucleic acid copy number was determined using PCR products of the target region as a template.
TextSentencer_T90 14339-14456 Sentence denotes The detection limit of nucleic acid copy number was determined using PCR products of the target region as a template.
TextSentencer_T91 14457-14554 Sentence denotes At least 10 2 copies/5 l of the template were required for detection by RT-LAMP (data not shown).
TextSentencer_T91 14457-14554 Sentence denotes At least 10 2 copies/5 l of the template were required for detection by RT-LAMP (data not shown).
TextSentencer_T92 14555-14683 Sentence denotes The RT-LAMP assay was carried out using a mixed sample of subgroups A and B RSV to evaluate potential interference between them.
TextSentencer_T92 14555-14683 Sentence denotes The RT-LAMP assay was carried out using a mixed sample of subgroups A and B RSV to evaluate potential interference between them.
TextSentencer_T93 14684-14824 Sentence denotes RT-LAMP was able to differentiate between RSV subgroups, and each primer set detected specifically its own target subgroup (data not shown).
TextSentencer_T93 14684-14824 Sentence denotes RT-LAMP was able to differentiate between RSV subgroups, and each primer set detected specifically its own target subgroup (data not shown).
TextSentencer_T94 14826-15157 Sentence denotes RT-LAMP assay was performed to evaluate whether the primer set used in this study could detect RSV in clinical specimens using 59 nasopharyngeal aspirates from children with respiratory tract infections, and the results were compared with those obtained using virus isolation and the EIA test, BD Directigen EZ RSV test (Table 4 ).
TextSentencer_T94 14826-15157 Sentence denotes RT-LAMP assay was performed to evaluate whether the primer set used in this study could detect RSV in clinical specimens using 59 nasopharyngeal aspirates from children with respiratory tract infections, and the results were compared with those obtained using virus isolation and the EIA test, BD Directigen EZ RSV test (Table 4 ).
TextSentencer_T95 15158-15277 Sentence denotes 61% (36/59) were positive by RT-LAMP, 34% (20/59) by viral isolation, 56% (26/46) by EZ RSV, and 58% (34/59) by RT-PCR.
TextSentencer_T95 15158-15277 Sentence denotes 61% (36/59) were positive by RT-LAMP, 34% (20/59) by viral isolation, 56% (26/46) by EZ RSV, and 58% (34/59) by RT-PCR.
TextSentencer_T96 15278-15385 Sentence denotes The success rate of RSV detection using the RT-LAMP assay was higher than virus isolation and the EIA test.
TextSentencer_T96 15278-15385 Sentence denotes The success rate of RSV detection using the RT-LAMP assay was higher than virus isolation and the EIA test.
TextSentencer_T97 15386-15617 Sentence denotes Of the 39 RSV isolation-negative samples, 4 had parainfluenza virus, 3 had influenza virus, and RSV isolation negative (n =2 6 ) 9 1 7 1 1 1 5 EZ RSV positive (n = 1 0 ) 9 1 9 1 EZ RSV negative (n = 16) 0 16 2 14 1 had enterovirus.
TextSentencer_T97 15386-15617 Sentence denotes Of the 39 RSV isolation-negative samples, 4 had parainfluenza virus, 3 had influenza virus, and RSV isolation negative (n =2 6 ) 9 1 7 1 1 1 5 EZ RSV positive (n = 1 0 ) 9 1 9 1 EZ RSV negative (n = 16) 0 16 2 14 1 had enterovirus.
TextSentencer_T98 15618-15839 Sentence denotes These other respiratory viruses isolated were RT-LAMP-negative except for one influenza virus-positive specimen that was RSV positive using the primer set described above as well as the primer set reported by Ushio et al.
TextSentencer_T98 15618-15839 Sentence denotes These other respiratory viruses isolated were RT-LAMP-negative except for one influenza virus-positive specimen that was RSV positive using the primer set described above as well as the primer set reported by Ushio et al.
TextSentencer_T99 15840-15978 Sentence denotes Comparison between the ability of EZ RSV, RSV isolation, and the RT-LAMP assay to detect RSV from clinical specimens is shown in Table 5 .
TextSentencer_T99 15840-15978 Sentence denotes Comparison between the ability of EZ RSV, RSV isolation, and the RT-LAMP assay to detect RSV from clinical specimens is shown in Table 5 .
TextSentencer_T100 15979-16101 Sentence denotes Of the 20 RSV isolation-positive specimens, 100% (20/20) were positive by RT-LAMP and 80% were positive (16/20) by EZ RSV.
TextSentencer_T100 15979-16101 Sentence denotes Of the 20 RSV isolation-positive specimens, 100% (20/20) were positive by RT-LAMP and 80% were positive (16/20) by EZ RSV.
TextSentencer_T101 16102-16208 Sentence denotes Of the 26 RSV isolation-negative specimens, 42% (11/26) were positive by RT-LAMP and 35% (9/26) by EZ RSV.
TextSentencer_T101 16102-16208 Sentence denotes Of the 26 RSV isolation-negative specimens, 42% (11/26) were positive by RT-LAMP and 35% (9/26) by EZ RSV.
TextSentencer_T102 16209-16262 Sentence denotes The sensitivity of RT-PCR was similar to that of EIA.
TextSentencer_T102 16209-16262 Sentence denotes The sensitivity of RT-PCR was similar to that of EIA.
TextSentencer_T103 16263-16385 Sentence denotes However, 12.5% (2/16) of specimens that were both antigen detection and virus isolation negative were positive by RT-LAMP.
TextSentencer_T103 16263-16385 Sentence denotes However, 12.5% (2/16) of specimens that were both antigen detection and virus isolation negative were positive by RT-LAMP.
TextSentencer_T104 16386-16515 Sentence denotes This paper describes a highly sensitive and specific genetic diagnostic method that can detect human RSV using the RT-LAMP assay.
TextSentencer_T104 16386-16515 Sentence denotes This paper describes a highly sensitive and specific genetic diagnostic method that can detect human RSV using the RT-LAMP assay.
TextSentencer_T105 16516-16636 Sentence denotes Subgroup A and B RSV were both detected by RT-LAMP using primer sets A or B, while other respiratory pathogens were not.
TextSentencer_T105 16516-16636 Sentence denotes Subgroup A and B RSV were both detected by RT-LAMP using primer sets A or B, while other respiratory pathogens were not.
TextSentencer_T106 16637-16721 Sentence denotes The assay developed in this study can detect viral RNA in 10 −1 to 10 −2 PFU of RSV.
TextSentencer_T106 16637-16721 Sentence denotes The assay developed in this study can detect viral RNA in 10 −1 to 10 −2 PFU of RSV.
TextSentencer_T107 16722-16938 Sentence denotes RSV was also detected in nasopharyngeal aspirates from children with respiratory tract infections, suggesting that the RSV RT-LAMP developed in the present study may be used for diagnosing RSV infections in children.
TextSentencer_T107 16722-16938 Sentence denotes RSV was also detected in nasopharyngeal aspirates from children with respiratory tract infections, suggesting that the RSV RT-LAMP developed in the present study may be used for diagnosing RSV infections in children.
TextSentencer_T108 16939-17002 Sentence denotes RSV infections are usually detected by virus isolation or EIAs.
TextSentencer_T108 16939-17002 Sentence denotes RSV infections are usually detected by virus isolation or EIAs.
TextSentencer_T109 17003-17080 Sentence denotes The virus isolation method, however, is only able to detect infectious virus.
TextSentencer_T109 17003-17080 Sentence denotes The virus isolation method, however, is only able to detect infectious virus.
TextSentencer_T110 17081-17195 Sentence denotes RSV is thermolabile and the quantity of infectious particles depends on sample preparation and storage conditions.
TextSentencer_T110 17081-17195 Sentence denotes RSV is thermolabile and the quantity of infectious particles depends on sample preparation and storage conditions.
TextSentencer_T111 17196-17320 Sentence denotes Thus, isolation of RSV can be difficult and RSV infection runs the risk of being overlooked if this is the only method used.
TextSentencer_T111 17196-17320 Sentence denotes Thus, isolation of RSV can be difficult and RSV infection runs the risk of being overlooked if this is the only method used.
TextSentencer_T112 17321-17575 Sentence denotes EIA kits do not require infectious viral antigens for detection and can be carried out easily in a short time (approximately 15 min), and are considered useful for general clinical diagnosis of young children (Ohm-Smith et al., 2004; Wren et al., 1990) .
TextSentencer_T112 17321-17575 Sentence denotes EIA kits do not require infectious viral antigens for detection and can be carried out easily in a short time (approximately 15 min), and are considered useful for general clinical diagnosis of young children (Ohm-Smith et al., 2004; Wren et al., 1990) .
TextSentencer_T113 17576-17692 Sentence denotes However, misdiagnosis can occur in 10-30% of RSV infections when the EIA kit is used alone (Grondahl et al., 2005) .
TextSentencer_T113 17576-17692 Sentence denotes However, misdiagnosis can occur in 10-30% of RSV infections when the EIA kit is used alone (Grondahl et al., 2005) .
TextSentencer_T114 17693-17934 Sentence denotes RSV is not detected easily in specimens from adult patients using either virus isolation or the EIA kit, because neutralizing antibodies maintain the virus titer low (Casiano-Colon et al., 2003; Falsey et al., 2002; Falsey and Walsh, 2000) .
TextSentencer_T114 17693-17934 Sentence denotes RSV is not detected easily in specimens from adult patients using either virus isolation or the EIA kit, because neutralizing antibodies maintain the virus titer low (Casiano-Colon et al., 2003; Falsey et al., 2002; Falsey and Walsh, 2000) .
TextSentencer_T115 17935-18069 Sentence denotes Molecular diagnostic methods can detect viruses as long as genomic sequences exist, regardless of the integrity of the virus particle.
TextSentencer_T115 17935-18069 Sentence denotes Molecular diagnostic methods can detect viruses as long as genomic sequences exist, regardless of the integrity of the virus particle.
TextSentencer_T116 18070-18165 Sentence denotes As a result, molecular diagnostics method should be undertaken for definitive diagnosis of RSV.
TextSentencer_T116 18070-18165 Sentence denotes As a result, molecular diagnostics method should be undertaken for definitive diagnosis of RSV.
TextSentencer_T117 18166-18436 Sentence denotes The LAMP assay is simpler and quicker than RT-PCR (Hong et al., 2004; Parida et al., 2004) , and results can be obtained by viewing fluorescence or determining turbidity values without opening the sample tubes, minimizing concerns about contaminating amplified products.
TextSentencer_T117 18166-18436 Sentence denotes The LAMP assay is simpler and quicker than RT-PCR (Hong et al., 2004; Parida et al., 2004) , and results can be obtained by viewing fluorescence or determining turbidity values without opening the sample tubes, minimizing concerns about contaminating amplified products.
TextSentencer_T118 18437-18577 Sentence denotes In addition, expensive fluorescent probes and detection instruments that are required for real-time PCR, are unnecessary for the LAMP assay.
TextSentencer_T118 18437-18577 Sentence denotes In addition, expensive fluorescent probes and detection instruments that are required for real-time PCR, are unnecessary for the LAMP assay.
TextSentencer_T119 18578-18749 Sentence denotes Therefore, the LAMP assay is useful for qualitative detection of RSV, and RT-LAMP can be used to amplify quantitatively the virus genome in real-time (Hong et al., 2004) .
TextSentencer_T119 18578-18749 Sentence denotes Therefore, the LAMP assay is useful for qualitative detection of RSV, and RT-LAMP can be used to amplify quantitatively the virus genome in real-time (Hong et al., 2004) .
TextSentencer_T120 18750-18835 Sentence denotes The RT-LAMP assay may be useful not only for diagnostic tests but also for prognosis.
TextSentencer_T120 18750-18835 Sentence denotes The RT-LAMP assay may be useful not only for diagnostic tests but also for prognosis.
TextSentencer_T121 18836-18986 Sentence denotes In this study, nasopharyngeal aspirates were collected from children with respiratory tract infections, and RT-LAMP was able to detect the RSV genome.
TextSentencer_T121 18836-18986 Sentence denotes In this study, nasopharyngeal aspirates were collected from children with respiratory tract infections, and RT-LAMP was able to detect the RSV genome.
TextSentencer_T122 18987-19268 Sentence denotes The RT-LAMP assay developed in this study also detected RSV genomes in samples that were negative by virus isolation and EIA, suggesting that RT-LAMP is a more sensitive assay for the detection of RSV antigens and is more useful in diagnosing RSV infections than classical methods.
TextSentencer_T122 18987-19268 Sentence denotes The RT-LAMP assay developed in this study also detected RSV genomes in samples that were negative by virus isolation and EIA, suggesting that RT-LAMP is a more sensitive assay for the detection of RSV antigens and is more useful in diagnosing RSV infections than classical methods.
TextSentencer_T123 19269-19396 Sentence denotes The RT-LAMP assay may also be used for diagnosing RSV infections in adults, although further studies are required to test this.
TextSentencer_T123 19269-19396 Sentence denotes The RT-LAMP assay may also be used for diagnosing RSV infections in adults, although further studies are required to test this.
TextSentencer_T124 19397-19501 Sentence denotes In this study, supernatants of nasopharyngeal aspirates were used to detect RSV using the RT-LAMP assay.
TextSentencer_T124 19397-19501 Sentence denotes In this study, supernatants of nasopharyngeal aspirates were used to detect RSV using the RT-LAMP assay.
TextSentencer_T125 19502-19649 Sentence denotes Since some paramyxovirus virions are cell associated (Wechsler et al., 1985) , this assay may be even more sensitive if the cells are not spun out.
TextSentencer_T125 19502-19649 Sentence denotes Since some paramyxovirus virions are cell associated (Wechsler et al., 1985) , this assay may be even more sensitive if the cells are not spun out.
TextSentencer_T126 19650-19854 Sentence denotes Of the clinical specimens, one was RT-LAMP positive using the RSV primer sets described above and the primer set described by Ushio et al. (2005) as well as being RSV isolation negative, and EIA negative.
TextSentencer_T126 19650-19854 Sentence denotes Of the clinical specimens, one was RT-LAMP positive using the RSV primer sets described above and the primer set described by Ushio et al. (2005) as well as being RSV isolation negative, and EIA negative.
TextSentencer_T127 19855-19989 Sentence denotes Viral isolation was influenza virus positive, however, so it is thought that this specimen may have been coinfected with both viruses.
TextSentencer_T127 19855-19989 Sentence denotes Viral isolation was influenza virus positive, however, so it is thought that this specimen may have been coinfected with both viruses.
TextSentencer_T128 19990-20100 Sentence denotes Perhaps the RSV gene was present in such small amounts that it could only be detected using the RT-LAMP assay.
TextSentencer_T128 19990-20100 Sentence denotes Perhaps the RSV gene was present in such small amounts that it could only be detected using the RT-LAMP assay.
TextSentencer_T129 20101-20256 Sentence denotes Recently, Ushio et al. (2005) developed a diagnostic method for detecting the RSV genome using RT-LAMP using the nucleoprotein region sequences as primers.
TextSentencer_T129 20101-20256 Sentence denotes Recently, Ushio et al. (2005) developed a diagnostic method for detecting the RSV genome using RT-LAMP using the nucleoprotein region sequences as primers.
TextSentencer_T130 20257-20368 Sentence denotes However, the specificity of this method of RSV detection was only tested against three influenza virus strains.
TextSentencer_T130 20257-20368 Sentence denotes However, the specificity of this method of RSV detection was only tested against three influenza virus strains.
TextSentencer_T131 20369-20609 Sentence denotes In contrast, we examined the specificity of the RT-LAMP technique against multiple respiratory pathogens, including metapneumovirus, influenza viruses, human coronaviruses, adenoviruses, and bacteria, and found that they were not amplified.
TextSentencer_T131 20369-20609 Sentence denotes In contrast, we examined the specificity of the RT-LAMP technique against multiple respiratory pathogens, including metapneumovirus, influenza viruses, human coronaviruses, adenoviruses, and bacteria, and found that they were not amplified.
TextSentencer_T132 20610-20769 Sentence denotes When clinical specimens were tested using the RT-LAMP primer set described above and that described by Ushio et al. (2005) RSV genome detection was comparable.
TextSentencer_T132 20610-20769 Sentence denotes When clinical specimens were tested using the RT-LAMP primer set described above and that described by Ushio et al. (2005) RSV genome detection was comparable.
TextSentencer_T133 20770-20863 Sentence denotes Thus, the sensitivity of our RT-LAMP assay described is equivalent to Ushio's method (2005) .
TextSentencer_T133 20770-20863 Sentence denotes Thus, the sensitivity of our RT-LAMP assay described is equivalent to Ushio's method (2005) .
TextSentencer_T134 20864-20995 Sentence denotes Since RSV is a negative strand RNA virus that develops frequent mutations, two or more primer sets may be preferable for diagnosis.
TextSentencer_T134 20864-20995 Sentence denotes Since RSV is a negative strand RNA virus that develops frequent mutations, two or more primer sets may be preferable for diagnosis.
TextSentencer_T135 20996-21121 Sentence denotes The RT-LAMP assay presented in this report should aid laboratory diagnosis and epidemiological surveillance of RSV infection.
TextSentencer_T135 20996-21121 Sentence denotes The RT-LAMP assay presented in this report should aid laboratory diagnosis and epidemiological surveillance of RSV infection.
TextSentencer_T136 21122-21243 Sentence denotes of Blood and Biologics at the National Institute of Infectious Disease for providing genomic DNA of respiratory bacteria.
TextSentencer_T136 21122-21243 Sentence denotes of Blood and Biologics at the National Institute of Infectious Disease for providing genomic DNA of respiratory bacteria.