人腺病毒-B55实时荧光定量PCR扩增方法的建立

详细信息    查看全文 | 推荐本文 |

英文篇名：Establishment of fluorescence quantitation PCR to amplify human adenovirus-B55 作者：董磊 ; 刘娟 ; 艾现印 ; 马红雨 ; 全首祯 ; 姜涛 英文作者：DONG Lei;LIU Juan;AI Xianyin;MA Hongyu;QUAN Shouzhen;JIANG Tao;Clinical Laboratory Center,the General Hospital of Peoples' Liberation Army Air Force;Department of Blood Transfusion,the General Hospital of Peoples' Liberation Army Air Force;Army Health Company 32143of Chinese People's Liberation Army;Beijing Institute of Microbiology and Epidemiology,State Key Laboratory of Pathogen and Biosecurity; 关键词：人腺病毒 ; 实时荧光定量聚合酶链反应 ; 快速检测 英文关键词：Human adenovirus;;Real-time fluorescence quantitation polymerase chain reaction;;Rapid detection 中文刊名：SHYY 英文刊名：Laboratory Medicine 机构：空军总医院临床检验中心;空军总医院输血科;中国人民解放军32143部队卫生连;军事科学院军事医学研究院微生物流行病研究所; 出版日期：2019-03-30 出版单位：检验医学 年：2019 期：v.34 基金：国家科技重大专项(2017ZX10305501);; 军队后勤科研重大项目(AWS16J020) 语种：中文; 页：SHYY201903017 页数：4 CN：03 ISSN：31-1915/R 分类号：79-82

摘要

目的构建人腺病毒(HAdV)-B55的实时荧光定量聚合酶链反应(PCR)扩增方法,并验证其检测性能。方法采用Beacon Designer 7软件设计针对HAdV-B55 Hexon基因序列的特异性引物,建立HAdV-B55的实时荧光定量PCR扩增方法,并评估其敏感性及特异性。结果建立了检测HAdV-B55的实时荧光定量PCR扩增方法,反应敏感性为0.08 PFU/反应体系,且特异性良好,与相关HAdV B组病毒均未检测到交叉反应。结论成功建立了针对我国HAdV-B55的特异性实时荧光定量PCR检测方法,为HAdV-B55的快速检测及防控提供了有力的支持。
        Objective To establish real-time fluorescence quantitation polymerase chain reaction(PCR)to amplify human adenovirus(HAdV)-B55,and to verify its performance. Methods The specific primers for the Hexon sequence of HAdV-B55 were designed with Beacon Designer 7 software,and the real-time ?uorescence quantitation PCR was established. The sensitivity and speci?city were evaluated. Results The speci?c primers for HAdV-B55 were obtained,and the real-time fluorescence quantitation PCR for HAdV-B55 had been established.The sensitivity was 0.08 PFU/reaction,and the specificity was good. There was no cross reaction with HAdVB Conclusions The real-time ?uorescence quantitation PCR to detect HAdV-B55 has been established,which lays the foundation for the rapid detection and prevention of HAdV-B55.

引文

[1]SMITH J G,WIETHOFF C M,STEWART P L,et al.Adenovirus[J].Curr Top Microbiol Immunol,2010,343:195-224.
    [2]KAJON A E,DICKSON L M,METZGAR D,et al.Outbreak of febrile respiratory illness associated with adenovirus 11a infection in a Singapore military training cAMP[J].J Clin Microbiol,2010,48(4):1438-1441.
    [3]CHMIELEWICZ B,BENZLER J,PAULI G,et al.Respiratory disease caused by a species B2 adenovirus in a military camp in Turkey[J].J Med Virol,2005,77(2):232-237.
    [4]LU Q B,TONG Y G,WO Y,et al.Epidemiology of human adenovirus and molecular characterization of human adenovirus 55 in China,2009-2012[J].Influenza Other Respir Viruses,2014,8(3):302-308.
    [5]LIU J,NIAN Q G,ZHANG Y,et al.In vitro characterization of human adenovirus type 55 in comparison with its parental adenoviruses,types 11 and 14[J].PLoSOne,2014,9(6):e100665.
    [6]ZHU Z,ZHANG Y,XU S,et al.Outbreak of acute respiratory disease in China caused by B2 species of adenovirus type 11[J].J Clin Microbiol,2009,47(3):697-703.
    [7]BOLOTIN S,ROBERTSONB A V,ESHAGHI A,et al.Development of a novel real-time reverse-transcriptase PCR method for the detection of H275Y positive influenza AH1N1 isolates[J].J Virol Methods,2009,158(1-2):190-194.
    [8]CARR M J,SAYRE N,DUFFY M.Rapid molecular detection of the H275Y oseltamivir resistance gene mutation in circulating influenza A(H1N1)viruses[J].J Virol Methods,2008,153(2):257-262.