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目的评价热对流PCR应用于口岸现场甲型流感病毒快速检测的效果。方法设计热对流PCR的引物和探针,用于甲型流感病毒的检测。用甲型和乙型流感病毒毒株和入境发热人员的鼻咽拭子样本验证热对流PCR的检测灵敏度、特异性及稳定性,并与市售商品化实时荧光定量PCR试剂比较检测甲型流感病毒的效果。结果通过对8株甲型流感病毒和1株乙型流感病毒毒株进行检测,热对流PCR对H1、H3及H5亚型的甲型流感病毒毒株的检测下限为0.001~0.005HAU,具有较好的检测广谱性,且不与乙型流感病毒发生交叉反应。对155份鼻咽拭子样本进行检测,和实时荧光定量PCR相比,检测灵敏度为94.74%,特异性为97.44%,且具有很好的检测稳定性。结论热对流PCR具有很高的检测灵敏度和特异性,可用于口岸现场的甲型流感病毒快速筛查。
Objective To evaluate the effectiveness of thermal convection PCR in the rapid detection of influenza A virus at the port. Methods Thermal convection PCR primers and probes were designed for the detection of influenza A virus. The detection sensitivity, specificity and stability of thermal convection PCR were tested on samples of nasopharyngeal swabs of influenza A and B strains and on arrival fever and tested for Influenza A by comparison with commercially available real-time PCR reagents The effect of the virus. Results By detecting 8 strains of influenza A and 1 strain of influenza B virus, the detection limit of influenza virus strains H1, H3 and H5 subtypes by heat convection PCR was 0.001 ~ 0.005HAU, Good detection of broad spectrum, and does not cross-react with influenza B virus. 155 samples of nasopharyngeal swabs were tested. Compared with real-time fluorescence quantitative PCR, the detection sensitivity was 94.74% and the specificity was 97.44%, and the detection stability was good. Conclusion Thermal convection PCR has high detection sensitivity and specificity and can be used for rapid screening of influenza A virus at the port.