猪的胸膜肺炎放线杆菌、多杀性巴氏杆菌和副猪嗜血杆菌复合PCR检测方法的建立  

Establishment of a Multiplex-PCR for Detecting Actinobacillus pleuropneumoniae,Pasteurella multocida and Haemophilus parasuis

作　　者：米丰泉[1] 陈小玲[2] 王翠敏[1] 赵玉军[1] 王凤强[1] 

机构地区：[1]沈阳农业大学畜牧兽医学院,辽宁沈阳110161 [2]北京市农林科学院畜牧兽医研究所,北京100089

出　　处：《动物医学进展》2005年第5期85-88,共4页Progress In Veterinary Medicine

摘　　要：在已经建立猪胸膜肺炎放线杆菌(APP)、猪多杀性巴氏杆菌(PM)、副猪嗜血杆菌(HPS)的单项PCR诊断方法的基础上,通过对扩增条件的优化,成功地建立了APP、PM、HPS的复合PCR实验室诊断方法,利用一次PCR反应,即可同时扩增出APP的342bp、PM的457 bp 和HPS 的821 bp 的特异性片段。该复合PCR 能同时检测到100 个每种细菌或50 pg 的APP 或HPS 的DNA和500 pg的PM的DNA。该三重复合PCR的敏感性同已报道的单PCR 一致。该方法的建立对临床上进行这3 种疾病的鉴别诊断和混合感染的检测都具有重要意义。A multiplex PCR detecting APP, PM and HPS was developed based on published single PCRs for these pathogens. The PCR reagents, annealing temperature and the PCR procedure were optimized. A 342 bp fragment of APP, a 457 bp of PM and a 821 bp of HPS can be simultaneously amplified in one reaction. The mPCR can detect minimum 100 cells of each species or 50 pg of APP and HPS DNA or 500pg of PM DNA in one reaction. The sensitivity of the mPCR is quite comparable with their reported uniplex PCRs. The assay is a potential tool for differentiation of the 3 pathogens and diagnosis of mixed infection by these bacteria.

关 键 词：猪胸膜肺炎放线杆菌 猪多杀性巴氏杆菌 副猪嗜血杆菌 复合PCR 

分 类 号：S852.61[农业科学—基础兽医学] S854.43[农业科学—兽医学]