PMC:7668588 / 8924-10155 JSONTXT

{"project":"LitCovid-PubTator","denotations":[{"id":"156","span":{"begin":660,"end":661},"obj":"Gene"},{"id":"157","span":{"begin":225,"end":228},"obj":"Gene"},{"id":"158","span":{"begin":670,"end":680},"obj":"Species"},{"id":"159","span":{"begin":323,"end":328},"obj":"Chemical"}],"attributes":[{"id":"A156","pred":"tao:has_database_id","subj":"156","obj":"Gene:43740570"},{"id":"A157","pred":"tao:has_database_id","subj":"157","obj":"Gene:4335"},{"id":"A158","pred":"tao:has_database_id","subj":"158","obj":"Tax:2697049"},{"id":"A159","pred":"tao:has_database_id","subj":"159","obj":"MESH:D014867"}],"namespaces":[{"prefix":"Tax","uri":"https://www.ncbi.nlm.nih.gov/taxonomy/"},{"prefix":"MESH","uri":"https://id.nlm.nih.gov/mesh/"},{"prefix":"Gene","uri":"https://www.ncbi.nlm.nih.gov/gene/"},{"prefix":"CVCL","uri":"https://web.expasy.org/cellosaurus/CVCL_"}],"text":"RT-qPCR was performed using the One Step PrimeScript III RT-qPCR Mix (Takara Bio) as reported previously [14]. The reaction mixture (total volume = 20 μL) contained 10 μL of 2× One Step PrimeScript III RT-qPCR Mix, 0.4 μL of ROX Reference Dye, 2 μL of 10× primer-probe mixture, 2 μL of RNA sample, and 5.6 μL of RNase-free water. Two different sets of primer-probe mixture were used in this study. The first set was developed by NIID (NIID-qPCR) and used with the final concentration of 500 nM, 700 nM, and 200 nM of the forward primer, reverse primer, and FAM-labeled probe, respectively [15]. The second primer-probe set was reported by WHO for detection of E gene of SARS-CoV-2 and SARS (WHO-qPCR), and used 400 nM of each primer and 200 nM of the probe [9]. The NIID-qPCR reaction was performed using the StepOnePlus instrument (Applied Biosystems) with a thermal cycle program of 52°C for 5 min, 95°C for 10 sec, followed by 45 cycles of 95°C for 5 sec, and 60°C for 30 sec WHO-qPCR was performed with the same program. Cut-off values were set at the threshold cycle (Ct) value of 40. To quantify viral RNA, a standard curve was generated with 10-fold serial dilutions of synthesized standard RNA of the qPCR target sequences."}

{"project":"LitCovid-sentences","denotations":[{"id":"T75","span":{"begin":0,"end":110},"obj":"Sentence"},{"id":"T76","span":{"begin":111,"end":329},"obj":"Sentence"},{"id":"T77","span":{"begin":330,"end":397},"obj":"Sentence"},{"id":"T78","span":{"begin":398,"end":594},"obj":"Sentence"},{"id":"T79","span":{"begin":595,"end":761},"obj":"Sentence"},{"id":"T80","span":{"begin":762,"end":1024},"obj":"Sentence"},{"id":"T81","span":{"begin":1025,"end":1089},"obj":"Sentence"},{"id":"T82","span":{"begin":1090,"end":1231},"obj":"Sentence"}],"namespaces":[{"prefix":"_base","uri":"http://pubannotation.org/ontology/tao.owl#"}],"text":"RT-qPCR was performed using the One Step PrimeScript III RT-qPCR Mix (Takara Bio) as reported previously [14]. The reaction mixture (total volume = 20 μL) contained 10 μL of 2× One Step PrimeScript III RT-qPCR Mix, 0.4 μL of ROX Reference Dye, 2 μL of 10× primer-probe mixture, 2 μL of RNA sample, and 5.6 μL of RNase-free water. Two different sets of primer-probe mixture were used in this study. The first set was developed by NIID (NIID-qPCR) and used with the final concentration of 500 nM, 700 nM, and 200 nM of the forward primer, reverse primer, and FAM-labeled probe, respectively [15]. The second primer-probe set was reported by WHO for detection of E gene of SARS-CoV-2 and SARS (WHO-qPCR), and used 400 nM of each primer and 200 nM of the probe [9]. The NIID-qPCR reaction was performed using the StepOnePlus instrument (Applied Biosystems) with a thermal cycle program of 52°C for 5 min, 95°C for 10 sec, followed by 45 cycles of 95°C for 5 sec, and 60°C for 30 sec WHO-qPCR was performed with the same program. Cut-off values were set at the threshold cycle (Ct) value of 40. To quantify viral RNA, a standard curve was generated with 10-fold serial dilutions of synthesized standard RNA of the qPCR target sequences."}