A culture-free method for rapidly and accurately quantifying active SARS-CoV-2

	A culture-free method for rapidly and accurately quantifying active SARS-CoV-2
	Yang, Y; Feng, XL; Pan, Y; Wang, X; Peng, T; Niu, CY; Qu, W; Zou, QC; Dong, LH; Dai, XH; Li, MH; Fang, X
	2023
发表期刊	ANALYTICAL AND BIOANALYTICAL CHEMISTRY
ISSN	1618-2642
卷号	415 期号:23 页码:5745-5753
摘要	Determining the quantity of active virus is the most important basis to judge the risk of virus infection, which usually relies on the virus median tissue culture infectious dose (TCID50) assay performed in a biosafety level 3 laboratory within 5-7 days. We have developed a culture-free method for rapid and accurate quantification of active severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) by targeting subgenomic RNA (sgRNA) based on reverse transcription digital PCR (RT-dPCR). The dynamic range of quantitative assays for sgRNA-N and sgRNA-E by RT-dPCR was investigated, and the result showed that the limits of detection (LoD) and quantification (LoQ) were 2 copies/reaction and 10 copies/reaction, respectively. The delta strain (NMDC60042793) of SARS-CoV-2 was cultured at an average titer of 10(6.13) TCID50/mL and used to evaluate the developed quantification method. Copy number concentrations of the cultured SARS-CoV-2 sgRNA and genomic RNA (gRNA) gave excellent linearity (R-2 = 0.9999) with SARS-CoV-2 titers in the range from 500 to 10(5) TCID50/mL. Validation of 63 positive clinical samples further proves that the quantification of sgRNA-N by RT-dPCR is more sensitive for active virus quantitative detection. It is notable that we can infer the active virus titer through quantification of SARS-CoV-2 sgRNA based on the linear relationship in a biosafety level 2 laboratory within 3 h. It can be used to timely and effectively identify infectious patients and reduce unnecessary isolation especially when a large number of COVID-19 infected people impose a burden on medical resources.
收录类别	SCI
语种	英语
文献类型	期刊论文
条目标识符	http://ir.kiz.ac.cn/handle/152453/14385
专题	管理、支撑部门_高等级生物安全灵长类动物实验中心
推荐引用方式 GB/T 7714	Yang, Y,Feng, XL,Pan, Y,et al. A culture-free method for rapidly and accurately quantifying active SARS-CoV-2[J]. ANALYTICAL AND BIOANALYTICAL CHEMISTRY,2023,415(23):5745-5753.
APA	Yang, Y.,Feng, XL.,Pan, Y.,Wang, X.,Peng, T.,...&Fang, X.(2023).A culture-free method for rapidly and accurately quantifying active SARS-CoV-2.ANALYTICAL AND BIOANALYTICAL CHEMISTRY,415(23),5745-5753.
MLA	Yang, Y,et al."A culture-free method for rapidly and accurately quantifying active SARS-CoV-2".ANALYTICAL AND BIOANALYTICAL CHEMISTRY 415.23(2023):5745-5753.

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