利用菌落多重PCR方法进行不同动物来源多杀性巴氏杆菌鉴定的研究

Identification of Pasteurella multocida strains isolated from different animals by the colony multiplex PCR

参照文献报道的多杀性巴氏杆菌种和荚膜型的特异基因合成6对特异引物,建立多杀性巴氏杆菌鉴定的菌落多重PCR方法,结果5株多杀性巴氏杆菌荚膜型参考菌株均扩增出了相应的预期片段,而支气管败血波氏杆菌、胸膜肺炎放线杆菌、大肠埃希菌的扩增均为阴性。利用此多重PCR方法对48株不同动物来源的多杀性巴氏杆菌进行了鉴定和荚膜型分型,同时与间接血凝试验以及Biolog细菌快速鉴定系统进行比对,结果表明,所建立的多重PCR方法与间接血凝试验、Biolog细菌快速鉴定系统的符合率均达到100%。

Six primers against Species-specific gene and capsular serogroup-specific genes were designed and the colony multiplex PCR of identification and serogroup of Pasteurella multocida were developed. Five Pasteurella multocida reference strains including A, B, D, E and F serotypes were identified using the colony multiplex PCR assay. The expected sequences were obtained successfully by the colony multiplex PCR assay. However, the sequences were not obtained from E. coli, B. bronchiseptica and A . pleuropneumoniae. Forty-eight Pasteurella multocida strains isolated from different animals were identified using the colony multiplex PCR assay, and the results were compared with Biolog automatic microbiology analysis system and Cater indirect hemagglutination test . The results showed good consistency. The coincidence rate between the results of the colony multiplex PCR assay and Cater indirect hemagglutination test and Biolog were all 100%.