期刊
INTERNATIONAL JOURNAL OF INFECTIOUS DISEASES
卷 102, 期 -, 页码 14-16出版社
ELSEVIER SCI LTD
DOI: 10.1016/j.ijid.2020.10.047
关键词
-
资金
- Universidad de Las Americas(Quito, Ecuador)
A new triplex RT-qPCR assay based on CDC primers and probes showed 100% specificity and 97.7% sensitivity, with a limit of detection at 1000 copies/mL. This assay could provide a reliable and efficient method for SARS-CoV-2 diagnosis, especially in times of testing supply shortages.
Background: Several RT-qPCR kits are available for SARS-CoV-2 diagnosis and some have emergency use authorization from the US Food and Drug Administration. In particular, the nCoV19 CDC kit includes two targets for detecting SARS-CoV-2 (N1 and N2) and an RNaseP (RP) target for RNA extraction quality control, all of which are labeled with FAM, and thus three PCR reactions are required per sample. Methods: We designed a triplex RT-qPCR assay based on nCoV19 primers and probes where N1, N2, and RP are labeled with FAM, HEX, and Cy5, respectively, so only a single PCR reaction is required for each sample for SARS-CoV-2 diagnosis. Results: In total, 172 samples were analyzed in both singleplex and triplex assays, where 86 samples tested SARS-CoV-2 negative with both assays, so the triplex assay specificity was 100%. In addition, 86 samples tested SARS-Co-V 2 positive with the singleplex assay and 84 with the triplex assay, so the sensitivity was 97.7%. The limit of detection for the triplex assay was determined as 1000 copies/mL. Conclusions: This new triplex RT-qPCR assay based on primers and probes from the CDC protocol is highly reliable for SARS-CoV-2 diagnosis, and it could speed up detection and save reagents during the current SARS-CoV-2 testing supplies shortage. (C) 2020 The Authors. Published by Elsevier Ltd on behalf of International Society for Infectious Diseases.
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