多重PCR在多杀性巴氏杆菌荚膜定群中的应用

Application of multiplex PCR for identification of Pasteurella multocida capsular sero-group

为了修订行业标准《猪巴氏杆菌病诊断技术》(NY/T 564-2002),改进多杀性巴氏杆菌荚膜定群方法,制备了多杀性巴氏杆菌分群抗血清,并建立了多重PCR,对53株菌同时用血凝方法及多重PCR进行定群。经正向间接血凝试验测定,制备的各型血清仅与对应型抗原发生凝集反应,表明血清具有较高的特异性,可作为多杀性巴氏杆菌的荚膜定群抗血清。53株多杀性巴氏杆菌均扩增出了相应的预期片段,PCR结果与Carter氏间接血细胞凝集试验结果相一致。结果表明,可将多重PCR增添至行业标准中,用其代替原有血清学方法进行多杀性巴氏杆菌荚膜定群。

In order to revise the industry standard Diagnostic Techniques of Swine Pasteurellosis(NY/T 564-2002),both the hemagglutination method with self-prepared anti-Pasteurella multocida antiserum and the colony multiplex PCR were used to identify 53 strains of P.multocida.The antiserum strictly agglutinated with corresponding antigen by Carter's indirect hemagglutination test,indicating that the serum was specific and might be used for grouping P.multocidacapsular.The PCR results were consistent with Carter's indirect hemagglutination test,showing the expected PCR products in these 53 strains.In conclusion,the multiplex PCR can be used as the industry standard instead of Carter's indirect hemagglutination test in identifying P.multocidacapsular sero-group.