作　　者： ; ; ; ; ; ; ; ; ; ; ; ; ;

机构地区： 深圳出入境检验检疫局

出　　处： 《中国预防兽医学报》 2013年第1期48-53,共6页

摘　　要： 自甲型H1N1(2009)流感病毒于2009年在世界多个国家暴发流行后,许多国家的猪群中检测到该病毒的存在,因此建立快速准确的甲型H1N1流感病毒的检测方法成为迫切需求。本研究针对甲型H1N1(2009)流感病毒NA基因设计特异性引物和探针,建立甲型H1N1(2009)流感病毒特异性实时荧光RT-PCR快速检测方法。并将该方法与WHO推荐美国CDC建立的检测方法和美国农业部推荐的检测方法进行比较。通过田间试验验证该方法在实际应用中的效果。结果表明:本研究建立的方法对检测甲型H1N1流感病毒裂解疫苗的灵敏度达到10-6,与WHO推荐的A型流感病毒实时荧光PCR检测方法的灵敏度一致,并且高于美国农业部推荐方法的检测灵敏度(10-5);该方法特异性强,与经典型H1N1和其他亚型流感病毒株无任何交叉反应。与香港兽医化验所进行了检测比对,检测灵敏度和特异性完全一致。近3 800份的田间试验表明,所建立的方法准确可靠。本研究所建立检测方法快速灵敏,检测结果准确可靠,适合用于甲型H1N1(2009)流感病毒的分子生物学检测和监测。 Pandemic H1N1 2009 influenza virus has been detected in pig herds on more than one continent since it outbroke in human in 2009. It is potential for this pandemic virus to resort or mutate in pigs. The rapid diagnosis is crucial to any control program. A TaqMan real-time RT-PCR （pH1N1 （2009）-rRT-PCR） test based on the NA gene of Pandemic H1N1 2009 was developed in the present study. The evaluation results indicated that the sensitivity of the pH1N1 （2009）-rRT-PCR was 106 which was equal to the CDC-InfA-rRT-PCR recommended by World Health Organization and superior to the USDA-pH1N1 （2009） -rRT-PCR recommended by Unite State Department Agriculture. The specificity test showed that the pH1N1 （2009）-rRT-PCR had no cross reactions with other virus. Comparing with Hongkong developed assay for 3,800 field sample detections indicated that the pH1N1 （2009）-rRT-PCR was a specific, sensitive and rapid assay, pH1N1 （2009）-rRT-PCR is an premising method for routinedetection of Pandemic H1N1 2009 and could be used to monitor and survey Pandemic H1N1 2009 influenza virus.

关 键 词： 甲型 流感病毒 实时荧光 快速检测

领　　域： [农业科学] [农业科学] [农业科学]