PCR检测方法在多杀性巴氏杆菌定种中的应用

Application of PCR for Identification of Pasteurella multocida

为修订行业标准《猪巴氏杆菌病诊断技术》(NY/T 564-2002),改进多杀性巴氏杆菌定种方法,采用16S r DNA基因序列分析法对中国兽医微生物菌种保藏管理中心(CVCC)保藏的63株猪源多杀性巴氏杆菌进行复核鉴定。63株猪源多杀性巴氏杆菌中有60株菌种与原鉴定结果一致。建立多杀性巴氏杆菌基于kmtΙ基因的PCR定种方法,对经16S r DNA PCR方法确认为多杀性巴氏杆菌的60株菌进行定种。结果表明,60株菌均扩增出了预期片段,基于kmtΙ基因的PCR定种方法与16S r DNA PCR方法试验结果相一致。研究结果显示,可将基于kmtΙ基因的PCR定种方法增添至行业标准中,作为原有生化鉴定方法的补充进行多杀性巴氏杆菌的定种。

In order to revise the industry standard Diagnostic Techniques of Swine Pasteurella （NY/T 564- 2002）, 16S rDNA sequence analysis was added to identify Pasteurella multocida. 63 strains of PasteureUa multocida from swine in China Veterinary Culture Collection Center（CVCC） were check identified with 16S rDNA sequence analysis. 60 strains of 63 strains suspected Pasteurella multocida were identified as Pasteurella multocida. Pasteurella multocida kmtI PCR identification method was constructed and applied to identify the above 60 strains of Pasteurella multocida. As a result, expected size DNA was amplified.The result is accordant with data of 16S rDNA sequence analysis. The result indicates that Pasteurella multocida kmtI PCR identification method can be added to the industry standard Diagnostic Techniques of Swine Pasteurella as Pasteurella multocida identification method besides original biochemical identification method