Optimization of a fluorescent qPCR detection for RNA of SARS-CoV-2.

Xuelong LI; Junhua Liu; Qianyang Liu; Lin YU; Shanshan WU; Xiushan Yin

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Optimization of a fluorescent qPCR detection for RNA of SARS-CoV-2.

Author: Xuelong LI ¹ ; Junhua LIU ¹ ; Qianyang LIU ² ; Lin YU ¹ ; Shanshan WU ¹ ; Xiushan YIN ¹
Author Information

1. School of Pharmaceutical and Biological Engineering, Shenyang University of Chemical Technology, Shenyang 110142, Liaoning, China.
2. BIOTECH (Shenyang) Biomedical Group Co., Ltd., Shenyang 110142, Liaoning, China.
Publication Type:Journal Article
Keywords: COVID-19; N gene; ORF1a gene; SARS-CoV-2; fluorescence real-time quantitative PCR
MeSH: Betacoronavirus; genetics; isolation & purification; Coronavirus Infections; diagnosis; virology; Early Diagnosis; Humans; Pandemics; Pneumonia, Viral; diagnosis; virology; Polymerase Chain Reaction; methods; standards; RNA, Viral; analysis; genetics; Sensitivity and Specificity; Time Factors
From: Chinese Journal of Biotechnology 2020;36(4):732-739
CountryChina
Language:Chinese
Abstract: We optimized a fluorescent quantitative polymerase chain reaction (qPCR) assay system for rapid and real time detection of SARS-CoV-2 RNA. The results show that the lowest dilution of RNA samples used for the detection of SARS-CoV-2 RNA could reach 1/10 000 (the initial value is set as 10 ng/μL). Moreover, the cycle threshold (Ct) for samples of clinically diagnosed COVID-19 was lower than 35 or 40. The sensitivity of this method was satisfactory. The results were consistent with those of the COVID-19 detection kit on the market under the same conditions, but the number of cycles required was shortened by about 2. Therefore, the optimized assay developed in this study can be used in screening and early clinical diagnosis. Our work provides a tool to facilitate rapid clinical diagnosis of COVID-19.