羊种布鲁氏菌疫苗株PCR-RFLP鉴定方法的建立

Development of a PCR-RFLP Method for Identification of Brucella melitensis Vaccine Strain Rev.1

为建立一种羊种布鲁氏菌Rev.1疫苗株PCR-RFLP鉴定方法,利用羊种布鲁氏菌Rev.1具有链霉素抗性的特性,选取与链霉素抗性相关编码核糖体蛋白S12的rps L基因为模板设计引物,经PCR扩增后,将产物进行Nci I酶切,发现羊种布鲁氏菌疫苗株Rev.1出现约500 bp条带,而不具有链霉素抗性的羊种布鲁氏菌株16M则出现384 bp条带。结果表明,PCR-RFLP方法能够用于羊种布鲁氏菌Rev.1疫苗株的鉴定,这为今后区分羊种布鲁氏菌疫苗株Rev.1免疫与野株感染提供了基础。

In order to develop a PCR-RFLP method for identifying the brucella melitensis vaccine strain Rev.1,given that the Rev.1 strain was resistant to streptomycin and the rps L gene was related to the streptomycin resistance,a pair of primers were designed according to the sequence of rps L gene in Gen Bank.After PCR amplification,the PCR products were digested with Nci I.Results showed that,a 500 bp band was got in the enzyme-digested products of the vaccine strain Rev.1,while a 384 bp band was got in the enzyme-digested products of wild strain 16 M.As a conclusion,the PCR-RFLP method could be applied in identification of the brucella melitensis vaccine strain Rev.1,which would provide a solid foundation for differentiating brucella melitensis vaccine strain Rev.1 and other wild strains in the future.