猪劳森氏胞内菌TaqMan荧光定量PCR检测方法的建立  

Establishment of TaqMan-based real-time PCR for detection of Lawsonia intracellularis

作　　者：陈世界[1] 杨苗[1] 林华[1] 郭杨[2] 严玉宝[1] 胡娟[1] 余华[1] 周岷江[1] 

机构地区：[1]四川出入境检验检疫局,四川成都610041 [2]四川省疾病预防控制中心,四川成都610041

出　　处：《中国预防兽医学报》2013年第7期570-573,共4页Chinese Journal of Preventive Veterinary Medicine

基　　金：国家质检总局课题(2009IK023);科技部质检公益性科研专项项目(200910188)

摘　　要：为建立一种快速有效的检测猪劳森氏胞内菌(LI)的方法,本研究根据该菌的天冬氨酸氨裂解酶基因保守序列设计合成一对特异性引物和一条TaqMan探针,建立了定量检测LI的荧光定量PCR方法,并对其进行敏感性、特异性、稳定性试验以及与套氏PCR方法的比较试验。结果表明,标准曲线的循环阈值与模板浓度呈现良好的线性关系,相关系数为0.998507;该方法对其他病原体的检测无特异性荧光信号;而且该方法灵敏度高于套式PCR方法。本研究为猪LI的检测提供了一种特异、敏感、快速的定量检测方法。In this study, a rapid and effective real-time PCR assay was developed for detection of Lawsonia intracellularis, with a pair of specific primers and a TaqMan probe designed according to the conserved sequence of L.intrncellularis aspartate ammonia-lyase gene in GenBank. The results indicated that standard curve showed a linear relationship between template concentration and threshold cycle, the correlation coefficient was 0.99. The method was demonstrated more sensitive than the nested PCR, and no cross-reaction was detected for other swine pathogens. The method could be used for a rapid and effective technique for the detection of L.intracellularis infection in pigs.

关 键 词：猪增生性肠病 劳森氏胞内菌 TAQMAN探针 实时荧光PCR 

分 类 号：S852.61[农业科学—基础兽医学]