荧光定量RT-PCR在流感病毒检测上的应用

Application of fluorescent real-time reverse transcriptase-polymerase chain reaction in detecting influenza viruses

目的 探讨实时荧光定量RT PCR检测方法在流行性感冒检测诊断上的意义。方法 收集集体暴发疑似流感、SARS患者的咽拭子标本 16起 2 0 7份和双份血清标本 10 4份 ,应用荧光定量RT PCR检测方法、MDCK细胞培养法和血凝抑制试验进行病原学和血清学检测。结果 2 0 7份集体暴发疑似流感、SARS患者的咽拭子标本中用荧光定量RT PCR检测 ,阳性 79份。阳性率 38 16 % (79 2 0 7)。MDCK细胞培养 ,16起中有 13起共 14 5份咽拭子有 6 2份标本阳性。阳性率 2 9 95 % (6 2 2 0 7)。两者的阳性率差异有显著意义 (χ2 =8.6 4 ,P <0 0 0 5 )。有 9起 10 4份成功的采集了双份血清 ,经HI试验有 6 4份双份血清与H3N2亚型抗原产生血抑 4倍增长 ,阳性率 6 1 5 4 % (6 4 10 4 )。结论 荧光定量RT PCR方法检测流感病毒能够在 3～ 4h内得出结果、且操作方便、特异性强、灵敏度高。因此我们认为实时荧光定量RT PCR方法 ,可作为一种快速的流感病毒检测诊断方法。

Objective To apply fluorescent real-time reverse transcriptase-polymerase chain reaction (RT-PCR) in detecting influenza viruses Methods A total of 207 oral swab samples were obtained in 16 collections from SARS patients and suspected influenza outbreak cases They were subjected to influenza virus detection by fluorescent real-time RT-PCR, MDCK cell culture, and hemagglutinin inhibition assay Results Out of 207 samples, 79 (3816%) were positive for influenza viruses when tested by fluorescent real-time PCR, and 62 (2995%) positive when tested by MDCK cell culture There was a statistically significant difference between them (χ 2=864, P<0005) From 104 cases in 9 collections dual serum samples were obtainable When tested with hemagglutinin inhibition assay, 64 cases (6154%) showed a 4-fold increase against H3N2 antigen Conclusion This study showed that fluorescent real-time PCR is a reliable, sensitive, and fast method for detecting influenza viruses