加温灭活对SARS-CoV-2咽拭子样本核酸检测结果的影响  

Influence of heating inactivation on the nucleic acid testing results of SARS-CoV-2 throat swab samples

作　　者：刘洁[1] 赵剑虹[1] 高艳[1] 韩桃利 李丽[1] 孔毅 赵焕兴 孙灵利[1] 王成彬 LIU Jie;ZHAO Jianhong;GAO Yan;HAN Taoli;LI Li;KONG Yi;ZHAO Huanxing;SUN Lingli;WANG Chengbin(Chaoyang Center for Disease Control and Prevention,Beijing 100021,China;Clinical Laboratory Center,the First Medical Center of Chinese People's Liberation Army General Hospital,Beijing 100853,China;The People's Liberation Army of China Troop 32382,Wuhan 432200,Hubei,China)

机构地区：[1]北京市朝阳区疾病预防控制中心,北京100021 [2]中国人民解放军总医院第一医学中心检验中心,北京100853 [3]中国人民解放军32382部队,湖北武汉432200

出　　处：《检验医学》2020年第5期405-408,共4页Laboratory Medicine

摘　　要：目的探讨实验室常用加温装置灭活升温时间及加温灭活对严重呼吸综合征冠状病毒2(SARSCoV-2)核酸检测结果的影响。方法收集SARS-CoV-2核酸阳性咽拭子样本10例,采用标准温度计标定样本的升温曲线,计算升温时间。将每例SARS-CoV-2核酸阳性咽拭子样本采样液各分装2份,一份56℃灭活30 min(56℃灭活处理组),另一份室温放置作为对照(未灭活处理组)。提取核酸后进行实时荧光定量聚合酶链反应(PCR)扩增。结果采用空气浴方式将样本升温至56℃需68 min,采用水浴方式需11.5 min。56℃灭活处理组核酸检测结果Ct值均增加;10份未经灭活处理的SARS-CoV-2核酸阳性样本中,56℃处理时有8份检测结果为阳性,加温灭活处理样本中出现假阴性结果与核酸浓度低有关。结论样本达到灭活温度所需时间与升温介质有关,升温灭活会影响SARS-CoV-2核酸检测效率。Objective To investigate the influence of heating inactivation on the nucleic acid testing results of severe acute respiratory syndrome coronavirus 2(SARS-CoV-2)throat swab samples.Methods Totally,10 throat swab samples of positive SARS-CoV-2 were collected.The standard thermometer was used to calibrate the temperature and calculate temperature rising time.Throat swab samples of positive SARS-CoV-2 were classified into 56℃30 min treatment group and untreatment group.Nucleic acid was extracted and amplified by real-time fluorescence quantitative polymerase chain reaction(PCR).Results It took 68 min to heat the sample to 56℃in air bath and 11.5 min in water bath.The results of PCR in 56℃30 min treatment group after heating showed that the Ct value increased.Of the 10 samples,8 samples were determined as positive in 56℃30 min treatment group.The samples with low nucleic acid concentration cannot be determined after treatment.Conclusions The heating up time depends on heating medium.Heating inactivation will affect SARS-CoV-2 nucleic acid testing results.

关 键 词：严重呼吸综合征冠状病毒2 灭活 温度 时间 核酸检测 

分 类 号：R446.1[医药卫生—诊断学]