应用多重聚合酶链反应检测四种霉形体的研究  

Study on detection of Mycoplasma gallisepticum, M.synoviae, M.iowae and M.meleagridis by multiplex polymerase chain reaction

作　　者：邓显文[1] 谢芝勋[1] 谢志勤[1] 廖敏[1] 庞耀珊[1] 刘加波[1] 唐小飞[1] 何竞铭[1] 

机构地区：[1]广西壮族自治区兽医研究所,广西南宁530001

出　　处：《中国兽医科技》2003年第6期14-17,共4页Chinese Journal of Veterinary Science and Technology

基　　金：广西留学基金资助项目 (桂科回 0 0 0 90 0 1)

摘　　要：根据鸡毒霉形体 (MG)、滑液囊霉形体 (MS)、衣阿华霉形体 (MI)和火鸡霉形体 (MM )的基因文库 ,设计了 4对分别与MG、MS、MI和MM某段基因序列互补的引物 ,用这 4对引物对同一样品中的MG、MS、MI和MM的DNA模板进行多重PCR扩增。结果 ,均同时得到了 4条特异性的大小与试验设计相符的 732bp(MG)、2 0 7bp(MS)、2 99bp(MI)、85 0bp(MM )多重PCR扩增带 ,而对其他 6种禽病病原体的PCR扩增结果均为阴性。敏感性测定结果表明 ,此多重PCR能同时检出 1pg的MG、MS。Four sets of specific primers were designed ac co rding to the sequences of Mycoplasma gallisepticum (MG), M.synoviae (MS) ,M.iowae(MI) and M.meleagridi s(MM) at the GenBank. A multiplex polymerse chain reaction was optimiz ed to simultaneously detect four pathogenic species of MG, MS, MI and MM. It wa s showed that all samples contained MG, MS, MI and MM DNA, can be amplified by the multiplex PCR using these four set primers, yielding four specific bands of 732 bp(MG),207 bp(MS),299 bp(MI) and 850 bp(MM). But no specific band was am plified from other six avian pathogenic viruses and bacteria. As little as 1 pg of MG, MS, MI and MM DNA can be detected using gel electrophoresis in this mul tiplex PCR.

关 键 词：火鸡霉形体 鸡毒霉形体 滑液囊霉形体 衣阿华霉形体 多重PCR 检测 鉴别 

分 类 号：S852.62[农业科学—基础兽医学]