人腺病毒-B55实时荧光定量PCR扩增方法的建立

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.