加热灭活对荧光定量逆转录PCR检测新型冠状病毒RNA影响分析  

Effect of Heat Inactivation on the Detection of the RNA of SARS-CoV-2 by RT-qPCR

作　　者：叶慧[1] 叶恒平[1] 马肖[1] 秦义组[2] 李涛 柴瑜[2] YE Hui;YE Hengping;MA Xiao;QIN Yizu;LI Tao;CHAI Yu(Xiantao Center for Disease Control and Prevention,Xiantao 433000,China;Anhui Provincial Center for Disease Control and Prevention,Hefei 230601,China)

机构地区：[1]湖北省仙桃市疾病预防控制中心,仙桃433000 [2]安徽省疾病预防控制中心,合肥230601

出　　处：《病毒学报》2020年第6期1004-1008,共5页Chinese Journal of Virology

摘　　要：56℃处理30min能有效灭活新型冠状病毒(SARS-CoV-2)。但加热处理对后继的荧光定量RT-PCR检测有无影响尚未见报道。本研究使用已知SARS-CoV-2核酸检测阳性的5个咽拭子标本,各自分为4份:对照组、56℃30min、56℃45min和56℃60min处理组。热处理后提取核酸并使用荧光定量RT-PCR检测病毒RNA相对含量。结果显示,56℃处理30~60min会导致荧光定量RT-PCR检测病毒核酸降低40%左右。56℃30min、56℃45min和56℃60min处理组间检测结果无统计学差异(P>0.05)。本研究提示,SARS-CoV-2标本56℃30min灭活后,再进行核酸提取和荧光定量RT-PCR检测,能更好地保护实验操作过程中人员和环境的安全。加热灭活对荧光定量RT-PCR检测结果的影响有限,但对于结果在临界值附近的标本应慎重使用。SARS-CoV-2 treated at 56℃for 30 min can be inactivated effectively.However,the effect of heat treatment on subsequent detection of the RNA of SARS-CoV-2 by real-time reverse transcription-quantitative polymerase chain reaction(RT-qPCR)has not been reported.We filled this knowledge gap in present study.We used five SARS-CoV-2-positive throat swabs.Each throat swab was divided into four parts and assigned to a group:control;56℃for 30 min;56℃for 45 min;56℃for 60 min.After heat treatment,SARS-CoV-2 RNA was extracted and detected by RT-qPCR(absolute quantitation using a standard curve).We found that SARSCoV-2 RNA was reduced by^40%after treatment at 56℃for 30–60 min.There was no significant difference(P>0.05 for all)in the test results between the treatment groups(56℃for 30 min;56℃for 45 min;56℃for60 min).Our study suggested that SARS-CoV-2 specimens could be inactivated at 56℃for 30 min,before RNA extraction and RT-qPCR detection,which could protect the safety of personnel and the environment during testing.Heat inactivation had a limited effect upon RT-qPCR detection but it should be used with caution if the specimen result is near the critical value.

关 键 词：新型冠状病毒(SARS-CoV-2) 生物安全 56℃灭活 RNA稳定性 荧光定量PCR(RT-qPCR) 

分 类 号：R373.1[医药卫生—病原生物学]