热对流PCR用于甲型流感病毒检测的评价

Abstract

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 efficiency of convective PCR on rapid detection of influenza A virus at frontier ports. Methods The convective PCR primers and probe were designed for the detection of influenza A virus.Influenza A virus strains,influenza B virus strain and nasopharyngeal swabs of entry travelers with fever were used to analyze the sensitivity,specificity and stability of convective PCR. The detection efficiency of convective PCR was compared with commercial Real-time PCR kit. Results A broad detection spectrum of convective PCR was observed in testing 8 influenza A virus strains with different genotypes（H1、H3 and H5）and 1 influenza B virus strain,the detection limit of convective PCR for influenza A virus strain was 0.001 ~0.005 HAU. There was no cross reaction with influenza B virus. When convective PCR was applied to the detection of 155 nasopharyngeal swabs,comparison with Real-time PCR,the detection sensitivity was 94.74%,the detection specificity was 97.44%,and the detection stability was good. Conclusion Convective PCR had a high sensitivity and specificity,was fit for the rapid detection for influenza A virus at frontier ports.