禽霍乱巴氏杆菌的分离鉴定及基因分型

发布时间：2018-11-20 15:57

【摘要】：为确定疑似禽霍乱病例病原种类及其病原基因型,本研究采用细菌分离技术对病原菌进行实验室分离培养,应用传统方法和分子生物学方法对分离细菌进行鉴定,并应用PCR扩增和基因序列分析对分离细菌进行基因分型。结果显示,分离菌具有多杀性巴氏杆菌(Pasteurella multocida,Pm)典型培养特征,菌落形态和菌体染色特征、生理生化特性均与多杀性巴氏杆菌相符;PCR扩增到457bp的基因片段。采用5对分型引物对分离菌进行基因分型显示,仅有A型引物扩增到大小1 050bp的目的基因片段,序列分析也显示分离菌荚膜基因与A型多杀性巴氏杆菌参考菌株荚膜特异性基因同源性高达97.6%~100.0%,系统进化与A型多杀性巴氏杆菌处于同一进化分支。结果表明,疑似禽霍乱病例病原为A型多杀性巴氏杆菌,本研究结果将为禽霍乱的防控提供参考资料。
[Abstract]:In order to identify the pathogenic species and their genotypes of suspected avian cholera cases, bacteria isolation techniques were used to isolate and culture pathogenic bacteria in laboratory, and traditional methods and molecular biological methods were used to identify the isolated bacteria. PCR amplification and gene sequence analysis were used for genotyping of isolated bacteria. The results showed that the isolated bacteria had typical culture characteristics of Pasteurella multocida (Pasteurella multocida,Pm), colony morphology, bacterial staining, physiological and biochemical characteristics were consistent with those of Pasteurella multocida, and 457bp gene fragment was amplified by PCR. Five pairs of primers were used for genotyping of isolated bacteria. Only A-type primers were used to amplify the target gene fragment with size of 1 050bp. The sequence analysis also showed that the homology of the capsule specific gene between the isolated strain and the reference strain of Pasteurella multocida was 97.6% and 100.0.The phylogeny was in the same evolutionary branch as that of Pasteurella multocida type A. The results showed that the suspected avian cholera case was caused by Pasteurella multocida type A. the results of this study will provide reference for the prevention and control of avian cholera.
【作者单位】：贵州大学动物科学学院;贵州省动物疫病与兽医公共卫生重点实验室;长顺县动物疫病预防控制中心;
【基金】：贵州省农业攻关项目“畜禽源主要病原菌药物防控技术创新与示范”(黔科合NY[2014]3042号) 贵州省科技创新人才团队项目(黔科合人才团队[2015]4016号)
【分类号】：S852.61

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