聚合酶螺旋反应快速检测甲型H1N1流感病毒

Rapid detection of influenza A/H1N1 virus by polymerase spiral reaction

目的建立利用聚合酶螺旋反应(PSR)快速检测甲型H1N1流感病毒的方法。方法针对甲型H1N1流感病毒的特异性HA基因设计了6套引物,通过实时浊度法和显色法两种方法判断结果。结果与结论从6套引物中筛选出了最佳引物,并确定最佳温度为65℃;进一步实验表明最佳引物能特异性地检测H1N1病毒,与14种其他呼吸道病原核酸无交叉反应,敏感性达到100拷贝,与PCR敏感性一致。所建立的方法简单快速、特异性强、敏感性高,适合现场和基层单位应用推广。

Objective To establish a polymerase spiral reaction（ PSR） method for rapid detection of influenza A/H1N1 virus. Methods Six sets of primers were designed for influenza A/H1N1 virus HA gene,and the results were determined with real time kinetic turbidimetric assay and colorimetry method. Results and Conclusion The best primers were selected from six sets of primers,and the best temperature was determined as 65 degrees Celsius. Further experiments showed that the best primer had good specificity for detection of influenza A/H1N1 virus,without cross reactions with 14 other respiratory tract pathogenic nucleic acids. The sensitivity was up to 100 copies,and consistent with that of PCR. So a PSR method is established for rapid detection of the influenza A/H1N1 virus,which is simple,quick,highly specific and sensitive,and especially applicable to field and grass-roots units.