摘要
目的探讨不同灭活条件对各种常见呼吸道病原体核酸检测结果的影响。方法收集2020年1月-2021年3月西藏军区总医院腺病毒、支原体、甲型流感病毒或乙型流感病毒核酸检测阳性病例鼻拭子和咽拭子样本12份,设置灭活温度梯度和时间梯度,分别于56、66、76和86℃处理30 min,于56℃分别处理30、60、120、240、480和960 min,比较不同条件灭活处理和未灭活处理样本实时荧光定量聚合酶链反应(qPCR)检测结果。结果对于腺病毒和支原体,56℃和66℃灭活处理30 min与未灭活处理样本的循环阈值(Ct)差异无统计学意义(P>0.05);76℃和86℃灭活处理30 min会使样本Ct值明显上升(P<0.05);56℃灭活30、60、120和240 min均不会造成样本Ct值的变化(P>0.05);56℃灭活480和960 min,会导致样本Ct值发生显著变化(P<0.05)。对于甲型流感病毒和乙型流感病毒,56℃灭活30 min与未灭活处理样本Ct值差异无统计学意义(P>0.05);66、76和86℃灭活30 min会使样本Ct值明显上升(P<0.05);56℃灭活30、60和120 min不会造成样本Ct值的变化(P>0.05);56℃灭活240、480和960 min会导致样本Ct值发生显著变化(P<0.05)。结论56℃灭活120 min以内不会改变病毒病原体qPCR检测结果。严重急性呼吸综合征冠状病毒2(SARS-CoV-2)的灭活处理不会对常见呼吸道病原体检测结果造成影响。
Objective To investigate the influence of different inactivating conditions on nucleic acid determination for respiratory pathogens.Methods A total of 12 patients who were positive of adenovirus,Mycoplasma,influenza A virus or influenza B virus in nasopharyngeal swabs were enrolled.Different inactivating temperatures and times were designed as follows:the samples were inactivated for 30 min in 56,66,76 and 86℃,as well as they were inactivated in 56℃for 30,60,120,240,480 and 960 min.The real-time fluorescence quantitative polymerase chain reaction(qPCR)results were compared between the groups before and after inactivation.Results For adenovirus and Mycoplasma,no statistical significance was found in cycling threshold(Ct)values among the groups that were inactivated for 30 min in 56 and 66℃and non-inactivation(P>0.05).An increase was determined in the groups that were inactivated for 30 min in 76 and 86℃(P<0.05).Inactivation in 56℃for 30,60,120 and 240 min did not result in changes in Ct values of samples(P>0.05).Inactivation in 56℃for 480 and 960 min would reduce the Ct values of samples(P<0.05).For influenza A virus and influenza B virus,no statistical significance was found in Ct values between the groups that were inactivated for 30 min in 56℃and non-inactivation(P>0.05).An increase was determined in the groups that were inactivated for 30 min in 66,76 and 86℃(P<0.05).Inactivation in 56℃for 30,60 and 120 min did not result in changes in Ct values of samples(P>0.05).Inactivation in 56℃for 240,480 and 960 min would reduce the Ct values of samples(P<0.05).Conclusions The samples inactivated in 56℃no more than 120 min would not influence the Ct values in qPCR.Inactivation for severe acute respiratory syndrome coronavirus 2(SARS-CoV-2)does not have effect on nucleic acid determination in other respiratory pathogens.
作者
王博文
曾晓华
彭凯歌
刘畅
武莹超
黄英眉
马盼盼
拉姆次仁
李国凯
杨莉
WANG Bowen;ZENG Xiaohua;PENG Kaige;LIU Chang;WU Yingchao;HUANG Yingmei;MA Panpan;Lamuciren;LI Guokai;YANG Li(Center for Disease Control and Prevention of Tibet Military Region,Lhasa 850000,Tibet,China;Department of Infectious Diseases,General Hospital of Tibet Military Region,Lhasa 850000,Tibet,China)
出处
《检验医学》
CAS
2021年第8期805-808,共4页
Laboratory Medicine
关键词
呼吸道病原体
灭活
温度
时间
实时荧光定量聚合酶链反应
Respiratory pathogen
Inactivation
Temperature
Time
Real-time fluorescence quantitative polymerase chain reaction
