猪繁殖与呼吸综合征RT-LAMP检测方法研究  

Development of RT-LAMP for Detecting Porcine Reproductive and Respiratory Syndrome Virus

作　　者：杨素[1] 沙才华[1] 顾海洋[2] 董尚智 廖秀云[1] 徐海聂[1] 薄清如[1] 罗宝正[1] 陈伟生[1] 

机构地区：[1]珠海出入境检验检疫局,广东珠海519015 [2]塔里木大学,新疆阿拉尔843300 [3]珠海市海洋农渔和水务局,广东珠海519000

出　　处：《中国畜牧兽医》2011年第6期136-141,共6页China Animal Husbandry & Veterinary Medicine

基　　金：珠海市科技计划项目(PC20081079)

摘　　要：应用反转录环介导等温扩增技术(RT-LAMP),利用猪繁殖与呼吸综合征病毒(PRRSV)核衣壳蛋白(N蛋白)ORF7基因保守序列的6个区域设计2对特异引物,经反应体系、反应条件优化及敏感性和特异性试验,建立了PRRSV RT-LAMP检测方法。研究结果表明,该检测方法不需要复杂仪器,仅用一台普通恒温水浴锅,在等温条件(63℃)保温1 h,即可完成核酸扩增反应,加入核酸染料SYBR GreenⅠ后,用肉眼即可对试验结果进行准确判定,为田间和基层部门检测猪繁殖与呼吸综合征病毒提供了一种廉价、简便、快速、敏感、特异的新方法。Applying reverse transcription LAMP（RT-LAMP）,according to the N protein conserved sequence of PRRSV ORF5 genes from six regions,two sets of primers were designed.By optimizing reaction system and conditions,the RT-LAMPmethod for detecting PRRSV was developed after sensitivity and specificity tests.The research results indicate that PRRSV nucleic acids amplification can be completed within 1 h in the isothermy condition of 63 ℃ only using a common constant temperature water-bath,test results can be judged with naked eyes after adding nucleic acids dye SYBR GreenⅠ.It provided a kind of cheap,simple,speedy,high sensitivity and specificity PRRSV detecting method for work field and grass-roots units.

关 键 词：猪繁殖与呼吸综合征病毒 RT-LAMP 检测方法 研究 

分 类 号：S852.65[农业科学—基础兽医学] Q78[农业科学—兽医学]