甲_3型流行性感冒病毒基因的快速检测

Rapid Detection of Influenza Type A_3 Virus

为建立流行性感冒 (流感 )的快速诊断技术 ,采用逆转录 聚合酶链反应 (RT PCR)方法检测甲3型流感病毒的血凝素 (HA)基因。对甲3型流感病毒的HA基因保守区域设计相关引物并进行PCR扩增 ,比较了它的特异性与灵敏度。结果显示 :该引物只能扩增甲3型流感病毒的特异性片段 ,而对其它型别的流感病毒以及麻疹、风疹、流行性腮腺炎病毒无交叉反应。检测的灵敏度 1次PCR可达 1 0TCID50 / 50ml以下 ,2次PCR反应可达 0 1TCID50 / 50ml以下。采用该方法从临床患者含漱液标本中的检出率 ,比采用狗肾传代细胞 (MDCK)分离病毒的阳性率更高 ,可达到迅速。

For establishing a method for rapid diagnosis of influenza,we detected the HA gene of human influenza A 3 virus by reverse transcription polymerease chain reaction(RT PCR) 1 5 pairs of primers corresponding to the conserved region of HA gene of A 3 influenza virus were designed,they can only amplify the specific segment of influenza A 3 virus,but not crossly react with measles,rubella and mumps viruses Sensitivity and specificity of the RT PCR method was compared to the conventional virus isolation method using MDCK cell The detection rate of A 3 influenza virus from patients' throat washing specimens by PCR method was much higher than that by conventional method The dilution experiments showed that influenza A 3 virus could be detected as few as 10TCID 50 /50ml in the first PCR and as few as 0 1TCID 50 /50ml in the second PCR Our results showed that PCR is a rapid,sensitive and reliable method for diagnosis of influenza A 3 infection