猪肺炎支原体和猪圆环病毒2型双重PCR检测方法的建立及应用  

Development of Duplex PCR Assay for Mycoplasma hypopneumoniae and Porcine Circovirus 2 Detection and Its Application

作　　者：胡军勇[1] 张倩[2,3] 王丹丹[2] 涂志勤[2] 胡睿铭[2] 汤细彪[2] 吴斌[2,3] 

机构地区：[1]华中农业大学动物科技学院,武汉430070 [2]华中农业大学动物传染病诊断中心,武汉430070 [3]华中农业大学动物医学院,武汉430070

出　　处：《畜牧兽医学报》2012年第11期1760-1766,共7页ACTA VETERINARIA ET ZOOTECHNICA SINICA

基　　金：湖北省研究与开发计划项目(2010BBB08)

摘　　要：本试验旨在建立一种针对猪圆环病毒2型(PCV2)和猪肺炎支原体(Mhp)的双重PCR检测方法。对猪伪狂犬病毒(PRV)、猪细小病毒(PPV)、沙门菌、大肠杆菌、猪链球菌、副猪嗜血杆菌、巴氏杆菌、支气管败血波氏杆菌基因组模板进行PCR特异性检测,没有任何非特异性扩增。敏感性试验显示,建立的PCR检测方法能够检测到的PCV2和Mhp模板的最低浓度分别为130和180fg.mL-1。同时用单项PCR和双重PCR对湖北省的各大猪场的174份PRDC样品进行检测,PCV2和Mhp的符合率分别达到100%和98.28%。进一步应用双重PCR调查PCV2和Mhp在不同猪场的感染动态,结果显示有一定比例的仔猪在产房就已经感染PCV2和Mhp,大部分猪是在保育和育肥阶段被感染;但是不同猪场表现出不同的感染动态。本试验建立了一种针对PCV2和Mhp的双重PCR检测方法,具有良好的特异性、敏感性,为临床疾病检测和疾病流行态势调查提供了有力的支持。A PCV2/Mhp duplex PCR detection assay was established in this paper. Specificity was tested by amplifying the genome DNA of PRV, PPV, Salmonella, E. coli, S. suis, Hps, Pm, B. bronchiseptica and none detectable amplification was found. Sensitivity of the duplex PCR assay proved that the minimal detectable template concentration were 130 and 180 fg·mL-1 for PCV2 and Mhp respectively. By applying both single and duplex PCR assays in PRDC cases in Hubei Province, 174 samples were tested, and the coincident rate between single and duplex PCR are 100% and 98.28% for PCV2 and Mhp respectively. By applying the duplex PCR assay, the infection dynamic of both PCV2 and Mhp were investigated in different herds, and the results suggested that the infection of both PCV2 and Mhp started in suckling stage with various proportions, and most infection took place in nursery and growing stages. Actually, different herds showed different infection dymanic. In this study, we established a duplex PCR assay and showed excellent specificity and sensitivity, which provides support for clinical diagnosis and epidemic investigation.

关 键 词：猪圆环病毒2型 猪肺炎支原体 PCR检测方法 感染动态 

分 类 号：S852.62[农业科学—基础兽医学] S852.659.2[农业科学—兽医学]