布鲁氏菌PCR鉴定方法的研究与应用

Study and application of polymerase chain reaction assay on Brucella identification

目的采用PCR、PCR-SSCP方法对布鲁氏菌进行快速鉴定,并对其种、型鉴别。方法分析已发表布鲁氏菌属、种、型基因序列,寻找出不同布鲁氏菌的种、型特异性碱基分布规律,设计特异性引物,用于布鲁氏菌属、种、型的鉴定;根据聚合酶链反应-单链构象多态性(PCR-SSCP)分析原理,建立PCR-SSCP方法,用于区分牛种A19疫苗株与其他型布鲁氏菌。结果所建立的PCR方法能高效、准确鉴定出布鲁氏菌,并能对其种、型进行区分;PCR-SSCP方法可将A19疫苗株从其他型布鲁氏菌中区分出来。结论利用PCR、PCR-SSCP方法能快速、准确地进行布鲁氏菌属、种、型以及疫苗株A19的鉴别,且简便、可靠,便于临床应用。

The purpose of this study was to apply PCR and PCR-SSCP assay method to identify Brucella and specific type of Brucella.In order to analyze a great number of partition Brucella of types sequence which have been issued as specific genotype and search for the distributing role of genotype-specific nucleotides,the specific primers were designed and PCR method was applied to identify specific type of Brucella.PCR-SSCP method was established to distinguish vaccine strain A19 from other specific types of Brucella.Results indicated that the established PCR method could be used to conduct the diagnosis of Brucellosis and identify specific type of Brucella.And PCR-SSCP assay could detect vaccine strain A19.It＇s suggested that PCR and PCR-SSCP assay is a reliable and convenient method to identify specific type of Brucella and vaccine strain A19.