Establishment of 2019-nCoV activity detection method based on PMA-RT-qPCR
10.3760/cma.j.cn112866-20210810-00140
- VernacularTitle:基于PMA-RT-qPCR的新型冠状病毒活性检测方法的建立
- Author:
Dandan LIU
1
;
Junhong LUO
;
Yiyue GE
;
Xiaojuan ZHU
;
Tao WU
;
Kangchen ZHAO
;
Xiangxi WANG
;
Fengcai ZHU
;
Baoli ZHU
;
Lunbiao CUI
Author Information
1. 南开大学药学院，天津 300000
- Keywords:
2019-nCoV;
Propidium monoazide;
RT-qPCR;
Active virus;
Inactivated virus
- From:
Chinese Journal of Experimental and Clinical Virology
2021;35(6):658-663
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To establish a rapid method for detecting the activity of 2019-nCoV2.Methods:The reverse transcription-quantitative polymerase chain reaction (RT-qPCR) detection system was screened after propidium monoazide (PMA) treatment and exposure to heat-inactivated 2019-nCoV samples, and the PMA pretreatment conditions were optimized to establish the 2019-nCoV PMA-RT-qPCR detection method . The established method was used to detect virus inactivated by different temperatures and chlorine-containing disinfectant, to evaluate its effect in detecting virus activity.Results:For the PMA-RT-qPCR assay, 200 μmol/L of PMA, 10 min of incubation time, 15 min of exposure time, and the CDC ORF1ab detection system were selected; there was no significant difference in the result of PMA-RT-qPCR and direct RT-qPCR for the active virus; the Ct values of PMA-RT-qPCR for virus inactivated by 95 ℃ and chlorine-containing disinfectant were higher than that of control groups at different dilutions; only partial dilutions of 70 ℃ and 56 ℃ heat-inactivated virus had higher Ct values than control groups. Conclusions:The established PMA-RT-qPCR for 2019-nCoV activity detection method has a good detection effect on the virus inactivated by 95 ℃ heat and chlorine disinfectant, and provides an auxiliary means for judging the infectivity of the virus in the sample.
