重组酶聚合酶扩增技术在人偏肺病毒快速诊断的方法建立与临床评价  

Establishment and Clinical Evaluation of Recombinase Polymerase Amplification for Rapid Diagnosis of Human Metapneumovirus

作　　者：唐娟[1] 刘文毅[1] 刘霄 TANG Juan;LIU Wenyi;LIU Xiao(Central Laboratory of Laboratory,Songgang People’s Hospital,Bao’an District,Shenzhen City,Guangdong Province 518105)

机构地区：[1]广东省深圳市宝安区松岗人民医院检验科中心实验室,518105

出　　处：《医学理论与实践》2021年第5期729-731,741,共4页The Journal of Medical Theory and Practice

基　　金：深圳市宝安区科技计划项目(20200518102810001)。

摘　　要：目的:研究重组酶聚合酶扩增技术在人偏肺病毒(hMPV)快速诊断的方法建立与临床价值。方法:将2017年1月—2019年12月医院收治的急性呼吸道感染患儿200例纳入研究。分离获取hMPV阳性病毒株,检索Genbank数据库,检索hMPV株基因序列,采用生物信息学软件通过靶向特定区域,对hMPV的N基因和L基因2个位点进行检测,并设计引物及荧光探针,使用病毒株和阴性对照品对基于重组酶聚合酶扩增技术快速检测hMPV的方法进行性能评估。同时,对所有受试者均进行直接荧光免疫法检测。以逆转录—聚合酶反应(RT-PCR)检测结果为金标准,对比不同检测方式诊断hMPV的效能。此外,分析hMPV感染患者合并其他病毒感染情况。结果:以RT-PCR检测结果为金标准,重组酶聚合酶扩增技术诊断hMPV的灵敏度、特异度、准确度均明显高于直接荧光免疫法检测(均P<0.05)。hMPV感染患者合并其他病毒感染发生率为26.83%,其中主要涵盖冠状病毒9.76%、呼吸道合胞病毒7.32%、流感病毒4.88%等。结论:重组酶聚合酶扩增技术在hMPV快速诊断中的应用效果较佳,可获得较为理想的灵敏度、特异度以及准确度,有望成为替代传统PCR的有效检测方式,值得临床推广应用。Objective:To study the establishment and clinical value of recombinase polymerase amplification technique in the rapid diagnosis of human metapneumovirus(hMPV).Methods:200 children with acute respiratory infection admitted to hospital from January 2017 to December 2019 were included in the study.Separation of hMPV positive strains,Genbank database search,retrieval hMPV plant gene sequences,such as using bioinformatics software by targeting specific areas,partial pulmonary virus to the person of L and N genes tested these two sites,and design primers and fluorescent probe,using strain and negative reference substance based on restructuring polymerase enzymes amplification technology rapid detection method for performance evaluation of hMPV.At the same time,all subjects were tested by direct fluorescent immunoassay.The results of reverse transcription-polymerase reaction(RT-PCR)were used as the gold standard to compare the diagnostic efficacy of different detection methods for hMPV.In addition,hMPV-infected patients with other virus infections were analyzed.Results:Using RT-PCR as the gold standard,the sensitivity,specificity and accuracy of recombinant polymerase amplification were significantly higher than detected by direct fluorescent immunoassay(all P<0.05).The incidence of hMPV infection with other viruses was 26.83%,including coronavirus 9.76%,respiratory syncytial virus 7.32%,influenza virus 4.88%,etc.Conclusion:Recombinase polymerase amplification technology has a good application effect in the rapid diagnosis of hMPV,and can obtain relatively ideal sensitivity,specificity and accuracy.It is expected to be an effective detection method instead of traditional PCR,and is worthy of clinical application.

关 键 词：人偏肺病毒 重组酶聚合酶扩增 逆转录—聚合酶反应 诊断效能 

分 类 号：R446[医药卫生—诊断学]