LitCovid-sentences  

PMC:7048229 / 3285-6090 JSONTXT 10 Projects

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Id Subject Object Predicate Lexical cue
T34 0-21 Sentence denotes Materials and methods
T35 23-40 Sentence denotes Sample collection
T36 41-282 Sentence denotes Human samples, including oral swabs, anal swabs and blood samples were collected by Wuhan pulmonary hospital with the consent from all patients and approved by the ethics committee of the designated hospital for emerging infectious diseases.
T37 283-317 Sentence denotes Two investigations were performed.
T38 318-422 Sentence denotes In the first investigation, we collected samples from 39 patients, 7 of which were in severe conditions.
T39 423-538 Sentence denotes In the second investigation, we collected samples from 139 patients, yet their clinical records were not available.
T40 539-608 Sentence denotes We only showed patients who were viral nucleotide detection positive.
T41 609-687 Sentence denotes Patients were sampled without gender or age preference unless where indicated.
T42 688-748 Sentence denotes For swabs, 1.5 ml DMEM+2% FBS medium was added in each tube.
T43 749-826 Sentence denotes Supernatant was collected after 2500 rpm, 60 s vortex and 15–30 min standing.
T44 827-896 Sentence denotes Supernatant from swabs were added to lysis buffer for RNA extraction.
T45 897-1054 Sentence denotes Serum was separated by centrifugation at 3000 g for 15 min within 24 h of collection, followed by 56°C 30 min inactivation, and then stored at 4°C until use.
T46 1056-1082 Sentence denotes RNA extraction and qRT-PCR
T47 1083-1182 Sentence denotes Whenever commercial kits were used, manufacturer’s instructions were followed without modification.
T48 1183-1265 Sentence denotes RNA was extracted from 200 μl of samples with the High Pure Viral RNA Kit (Roche).
T49 1266-1344 Sentence denotes RNA was eluted in 50 μl of elution buffer and used as the template for RT–PCR.
T50 1345-1432 Sentence denotes QPCR detection method based on 2019-nCoV S gene can be found in the previous study [5].
T51 1433-1556 Sentence denotes In brief, RNA extracted from above used in qPCR by HiScript® II One Step qRT-PCR SYBR® Green Kit (Vazyme Biotech Co., Ltd).
T52 1557-1751 Sentence denotes The 20 μl qPCR reaction mix contained 10 μl 2× One Step SYBR Green Mix, 1 μl One Step SYBR Green Enzyme Mix, 0.4 μl 50 × ROX Reference Dye 1, 0.4 μl of each primer (10 μM) and 2 μl template RNA.
T53 1752-1791 Sentence denotes Amplification was performed as follows:
T54 1792-1944 Sentence denotes 50°C for 3 min, 95°C for 30 s followed by 40 cycles consisting of 95°C for 10 s, 60°C for 30 s, and a default melting curve step in an ABI 7500 machine.
T55 1946-1962 Sentence denotes Serological test
T56 1963-2149 Sentence denotes In-house anti-SARSr-CoV IgG and IgM ELISA kits were developed using SARSr-CoV Rp3 NP as antigen, which shared above 90% amino acid identity to all SARSr-CoVs, as reported previously [5].
T57 2150-2262 Sentence denotes For IgG test, MaxiSorp Nunc-immuno 96 well ELISA plates were coated (100 ng/well) overnight with recombinant NP.
T58 2263-2317 Sentence denotes Human sera were used at 1:20 dilution for 1 h at 37°C.
T59 2318-2444 Sentence denotes An anti-Human IgG-HRP conjugated monoclonal antibody (Kyab Biotech Co., Ltd, Wuhan, China) was used at a dilution of 1:40,000.
T60 2445-2483 Sentence denotes The OD value (450–630) was calculated.
T61 2484-2605 Sentence denotes For IgM test, MaxiSorp Nunc-immuno 96 wellELISA plates were coated (500 ng/well) overnight with anti-human IgM (µ chain).
T62 2606-2766 Sentence denotes Human sera were used at 1:100 dilution for 40 min at 37°C, followed by anti-Rp3 NP-HRP conjugated (Kyab Biotech Co., Ltd, Wuhan, China) at a dilution of 1:4000.
T63 2767-2805 Sentence denotes The OD value (450–630) was calculated.