猪繁殖与呼吸综合征病毒美洲型RT-LAMP检测方法建立  

Development of an RT-LAMP Assay for Detection of American Type Porcine Reproductive and Respiratory Syndrome Virus(PRRSV)

作　　者：蒲静[1] 汪琳[1] 尹羿[1] 柏亚铎[1] 任彤[1] 张伟[1] 高志强[1] 乌日古玛拉[1] 时培蛟 肖春芳[1] 

机构地区：[1]北京出入境检验检疫局,北京100113

出　　处：《中国动物检疫》2015年第2期57-62,共6页China Animal Health Inspection

基　　金：国家质检总局科技计划项目(2013IK027)资助

摘　　要：本研究针对猪繁殖与呼吸综合征病毒美洲型ORF6基因的6个区域,利用2对引物建立检测猪繁殖与呼吸综合征病毒的RT-LAMP快速检测方法,该方法具有高灵敏度,高特异性的优点,在2小时左右即可得出结果,其灵敏度与PRRSV荧光RT-PCR检测方法相近,高于普通RT-PCR检测方法。将灭活的猪繁殖与呼吸综合征病毒美洲型培养物作10倍系列稀释,结果显示建立的荧光RT-PCR的检测极限可达10-7,RT-PCR检测方法的检测极限为10-5,RT-LAMP检测方法达到10-8,表明建立的RT-LAMP检测方法非常灵敏,可用于检测猪组织样本中的美洲型猪繁殖与呼吸综合征病毒。Two pairs of primers targeting six regions of American-type porcine reproductive and respiratory syndrome virus ORF6 gene, and RT-LAMP assay was established for the rapid detection of porcine reproductive and respiratory virus with high sensitivity and specificity. It could be finished in about 2 hours. The sensitivity of the assay was similar to PRRSV fluorescent RT-PCR, and higher than the normal RT-PCR. The inactivated porcine reproductive and respiratory syndrome virus culture was serially diluted in 10^-folds, and tested with PRRSV fluorescent RT-PCR, RT- LAMP assay and normal RT-PCR. The result showed that the fluorescence RT-PCR detection limit was up to 10^-7, the normal RT-PCR detection limit was 10^-5, and the RT-LAMP detection limit was 10^-8, indicating RT-LAMP assay was very sensitive. The assay could be used for rapid detection of the American-type porcine reproductive and respiratory syndrome virus in various pig tissue samples.

关 键 词：猪繁殖与呼吸综合征病毒 美洲型 RT-LAMP 

分 类 号：S852.651[农业科学—基础兽医学]