摘要
目的通过对1种新型呼吸道病原体核酸多重联检试剂盒检测效能评价,为新型冠状病毒(SARS-CoV-2)、流行性感冒(流感)病毒等叠加流行的大规模检测提供适宜技术方法。方法收集北京地区急性呼吸道感染病例阳性标本252份,采用4种核酸检测试剂盒(3种核酸多重检测试剂盒A、B、C及1种SARS-CoV-2检测试剂盒D)分别进行荧光聚合酶链式反应(PCR)检测,评价其与临床标本检测结果的一致性。选取3份呼吸道病原体标本核酸,使用无核糖核酸(RNA)酶水10倍梯度稀释,并选取1份SARS-CoV-2阳性标本核酸稀释为10^(−1)~10^(−6),评价A试剂盒的检出能力及批内重复性。结果A、B、C试剂盒对甲型流感病毒、鼻病毒、腺病毒、肠道病毒、人偏肺病毒、副流感病毒3型、副流感病毒4型、冠状病毒NL63的检出率均为100%,Kappa值均为1,结果完全一致。对乙型流感病毒、呼吸道合胞病毒、副流感病毒1型、副流感病毒2型、冠状病毒OC43、冠状病毒229E、冠状病毒HKU1及肺炎支原体的Kappa值范围为0.91~1。3种核酸联检试剂盒对呼吸道病原体的最低检出限相当。A与D试剂盒对SARS-CoV-2的阴阳性符合率均为100%,批内重复性显示在稀释度10^(−1)~10^(−3)范围内,2种试剂各重复检测5次变异系数(CV)值均<5%。在稀释度10^(−4)~10^(−5)范围内,A试剂对ORF1ab基因检出率为40%,N基因检出率为80%,D试剂对ORF1ab基因检出率为30%,N基因检出率为20%。结论A试剂盒与其他3种试剂盒对病毒载量较高的阳性标本检测的准确度无明显差异,检测性能相似;本研究为大规模核酸检测提供了检测试剂盒的多种选择。
Objective To evaluate the detection consistency and power of a multiplex combined real-time PCR detection kits,and provide reference for the prevention and control of influenza plus SARS-CoV-2 infection.Methods A total of 252 acute respiratory infectious samples were collected in Beijing.Four nucleic acid detection kits(A,B,C for multi nucleic acids and D for nucleic acid of SARS-CoV-2)were used for real time PCR,respectively,to evaluate the consistency of the test results.Three nucleic acid samples of respiratory pathogens in 10-fold dilutions with RNase-free water and 1 nucleic acid samples of SARS-CoV-2 in dilution of 10^(–1)–10^(–6) folds were used to evaluate the detection power and intra-batch repeatability of kit A.Results The positive coincidence rates of kit A,B and C to detect influenza A virus,rhinovirus,adenovirus,enterovirus,human metapneumovirus,parainfluenza virus type 3 and 4 and coronavirus NL63 were 100%(all Kappa values=1).The detection power of three assays for influenza B virus,respiratory syncytial virus,parainfluenza virus type 1 and 2,coronavirus OC43,229E,HKU1 were similar,with the overall consistency rates of 99.17%–100.00%(all Kappa values≥0.91).The detection power of kit A,B and C were similar(P>0.05).The SARS-CoV-2 negative and positive coincidence rates tested by kit A and kit D were 100%,the intra-batch repeatability showed the coefficient of variatiot(CV)of viral loads tested by these two kits were less than 5%in the dilution range of 10^(−1)–10^(−3).In the dilution range of 10^(−4)–10^(−5),the detection power of kit A for open reading frame(ORF)1ab gene was higher(40%)than kit D(30%),as well as for N gene(80%vs.20%).Conclusion There were no significant differences in the accuracy of the kit A and other three kits for positive samples with high viral loads,and the detection power were similar.This study provided a multiple choice for large-scale nucleic acid reference for the prevention and control of SARS-CoV-2 infection.
作者
罗琴
谢会
罗明
李爱华
黄琪
龚成
王雪
李茂中
Luo Qin;Xie Hui;Luo Ming;Li Aihua;Huang Qi;Gong Cheng;Wang Xue;Li Maozhong(School of Public Health,Capital Medical University,Beijing 100069,China;Beijing Center for Disease Control and Prevention/Beijing Academy for Preventive Medicine/Beijing Institute of Tuberculosis Control Research and Prevention,Beijing 100013,China)
出处
《疾病监测》
CAS
CSCD
北大核心
2022年第1期132-138,共7页
Disease Surveillance
基金
首都卫生发展科研专项(No.2020–4–3014)
科技部传染病重大专项(No.2017ZX10103004–002)
中共北京市委组织部(No.2018000021469G299)。
关键词
呼吸道病原体
新型冠状病毒
核酸检测
Respiratory pathogen
SARS-CoV-2
Detection of nucleic acids
