猪细环病毒LAMP检测方法的建立  

Development of loop-mediated isothermal amplification for detection of Torque Teno sus virus

作　　者：向海洋[1,2] 聂福平[2,3,4] 杨俊[2,3] 王昱[2,3,4] 肖进文[2,3] 李应国[2,3] 黄鹤[1,2] 聂奎[1] 

机构地区：[1]西南大学动物科技学院,重庆400715 [2]重庆出入境检验检疫局,重庆400020 [3]重庆市进出口食品安全工程中心,重庆400020 [4]重庆大学生物工程学院,重庆400030

出　　处：《中国预防兽医学报》2013年第9期747-751,共5页Chinese Journal of Preventive Veterinary Medicine

基　　金：国家质检总局科技项目(2011IK023);重庆市科委科技创新能力建设项目(CSTC;2009CB1012)

摘　　要：为建立一种特异和快速的猪细环病毒(TTSuV)检测方法,本研究通过比对TTSuV的全基因序列,选择其保守区域,设计了针对TTSuV检测的LAMP引物,利用LAMP Real Time Turbidimeter LA-320仪对反应体系及条件进行了优化,建立TTSuV环介导等温扩增的检测方法。结果表明:该方法最佳反应条件为64℃恒温50 min,病毒的最低检出限为5.5拷贝/μL,并且与其他相关传染病无交叉反应。临床样品检测结果显示,猪繁殖与呼吸综合征病毒(PRRSV)和2型猪圆环病毒(PCV2)阳性的样品中TTSuV呈高阳性率,分别为92%和87.3%,显著高于非PRRSV和PCV2阳性的猪群。该方法的建立为快速及特异性的检测猪TTSuV提供了有效的方法。To develop a loop-mediated isothermal amplification （LAMP） method for detection of porcine Torque Teno sus virus （TTSuV）, a set of primers for detecting TTSuV was designed according to conserved sequence of TTSuV. With the use of LAMP Real Time Turbidimeter LA-320, the LAMP method was developed under the optimized reaction conditions including the amplification in a constant temperature at 64 ℃ for 50 min. The results show that the method was specific for detection of TTSuV with a minimum detection of 5.5 copies/μL viral nucleic acid, and no cross-reactions with other related virus or bactera. In addition, clinical sample test results indicated that the positive rate of the TTSuV was higher in the PRRSV and PCV2 positive samples than that in PRRSV and PCV2 negative samples, and the TTSuV positive rates were 92% and 87.3%, respectively. Therefore, the rapid and easy-to-performed LAMP facilitates the detection of TTSuV for both laboratory and clinical diagnosis practice.

关 键 词：猪细环病毒 环介导等温扩增 检测 

分 类 号：S852.65[农业科学—基础兽医学]