多位点可变数目串联重复序列分型方法检测布鲁氏菌分型标准化操作方法的建立和应用

Establishment of standard operation procedures for multiple locus variable number tandem repeats analysis on typing of Brucella

摘要: 目的建立常用布鲁氏菌多位点可变数目串联重复序列分析(MLVA)的标准化方法及应用研究。方法选取16个位点,通过核酸提取、聚合酶链反应(PCR)、琼脂糖凝胶电泳、毛细管电泳等技术,结合BioNumerics(Version 5.0)软件,对16株布鲁氏菌标准菌株和不同地区分离的32株菌株进行分型。结果利用所建立的MLVA方法可以区分16株布鲁氏菌标准菌株,并可以将32株地方株区分为牛种和羊种两大类。结论初步建立了MLVA标准化方法,可以在种的水平鉴定布鲁氏菌,还可以追溯传染源,确定流行趋势。

Abstract: Objective To establish a standard operation procedures for multiple locus variable number tandem repeats analysis(MLVA)on subtyping of Brucella, and applied study. Methods Sixteen loci were selected,the subtyping of 16 standard strains of Brucella and 32 Brucella strains isolated from different areas were conducted by nucleic acid extraction, PCR, agarose gel electrophoresis, capillary electrophoresis and BioNumerics(version 5.0)software. Results The MLVA established could identify 16 standard strains of Brucella and classify 32 Brucella strains from different areas as Br melitensis and Br abortus. Conclusion The standard MLVA established can be used to identify Brucella species, trace the source of infection and determine the epidemic trend.