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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
引用本文:米丰泉,陈小玲,王翠敏,赵玉军,王凤强.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[J].动物医学进展,2005,26(5):85-88.
作者姓名:米丰泉  陈小玲  王翠敏  赵玉军  王凤强
作者单位:1. 沈阳农业大学畜牧兽医学院,辽宁,沈阳,110161;北京市农林科学院畜牧兽医研究所,北京,100089
2. 北京市农林科学院畜牧兽医研究所,北京,100089
3. 沈阳农业大学畜牧兽医学院,辽宁,沈阳,110161
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457bp和HPS的821bp的特异性片段。该复合PCR能同时检测到100个每种细菌或50pg的APP或HPS的DNA和500pg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。

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
文章编号:1007-5038(2005)05-0085-04
修稿时间:2004年12月10

Establishment of a Multiplex-PCR for Detecting Actinobacillus pleuropneumoniae,Pasteurella multocida and Haemophilus parasuis
MI Feng-quan,CHEN Xiao-ling,WANG Cui-min,ZHAO Yu-jun,WANG Feng-qiang.Establishment of a Multiplex-PCR for Detecting Actinobacillus pleuropneumoniae,Pasteurella multocida and Haemophilus parasuis[J].Progress In Veterinary Medicine,2005,26(5):85-88.
Authors:MI Feng-quan  CHEN Xiao-ling  WANG Cui-min  ZHAO Yu-jun  WANG Feng-qiang
Abstract: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.
Keywords:Actinobacillus pleuropneumoniae  Pasteurella multocida  Haemophilus parasuis  multiplex PCR
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