Application evaluation of a rapid fluorescence quantitative PCR method for the detection of SARS-CoV-2
10.3760/cma.j.cn112309-20210402-00107
- VernacularTitle:一种新型冠状病毒荧光定量PCR快速检测方法的应用评价
- Author:
Peihua NIU
1
;
Yaowu ZHU
;
Roujian LU
;
Jing PENG
;
Na ZHU
;
Yanjun LU
;
Wenling WANG
;
Ming NI
;
Wenjie TAN
Author Information
1. 中国疾病预防控制中心病毒病预防控制所应急技术中心，北京 102206
- Keywords:
SARS-CoV-2;
Real-time fluorescent quantitative PCR;
Point-of-care testing (POCT);
Detection limit;
Application evaluation
- From:
Chinese Journal of Microbiology and Immunology
2021;41(8):588-591
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To establish and evaluate a rapid nucleic acid detection method for SARS-CoV-2 based on COYOTE ? Flash20 real-time fluorescent quantitative PCR instrument. Methods:A rapid reaction system was constructed by using specific primer and probe sets targeting ORF1ab and N gene of SARS-CoV-2, and the sensitivity and specificity of the system were verified. At the same time, 108 clinical samples of COVID-19 were used to evaluate the application of this method.Results:The detection method did not require nucleic acid extraction, and the manual operation time was only one minute. After the sample was sent to the system, the test could be completed in 30 minutes. The detection limit of this method was 4×10 2 copies/ml. It had no cross-reactivity with other human coronaviruses (including HCoV-229E, HCoV-NL63, HCoV-OC43, HCoV-HKU1, SARS-CoV and MERS-CoV) and other respiratory viruses. The evaluation of clinical sample application showed that the total coincidence rate with the conventional RT-qPCR which required nucleic acid extraction was 98.15%. Conclusions:Through the application evaluation of the rapid fluorescent quantitative PCR method of SARS-CoV-2, it was found that the method was simple, fast, specific and sensitive, and it was suitable for real-time and rapid detection needs in varieties of situations.
