布氏杆菌分子亚型分析方法的建立与应用研究

Study on the development and application of molecular subtyping method for Brucella

目的应用高变八聚体寡核苷酸指纹(hypervariable octameric oligonucleotide finger_prints,HOOF_Prints)分析法对我国的布氏杆菌进行亚型和分子流行病学的研究。方法根据布氏杆菌8个数目可变串联重复单元(variable number of tandem repeats,VNTR)位点序列设计8对特异性的引物,用PCR方法扩增两株临床分离株并测序,得到8个位点的重复次数,进一步与国际上已公布的布氏杆菌VNTR数据进行比对及构建基因树,从而建立布氏杆菌亚型的分型方法。结果通过基因树得到我院两例布氏杆菌,均属猪二型,与我国流行的牛种与羊种布氏杆菌菌型存在基因型上的差异。结论HOOF_Prints分析法可在分子水平分析我国布氏杆菌的流行特点。

Objective To study the molecular epidemiological profiles of Brucella infection in China, the method of hypervariable octameric oligonucleotide finger-prints（HOOF-Prints） analysis was established. Methods We designed eight specific primers according to the sequences of Brucella＇s eight VNTR（ variable number of tandem repeats）loci and developed a PCR-based method to identify the number of repeat units at each locus. HOOF-Prints data of domestic isolate of Brucella was compared with international published VNTR genotypes data by constructing HOOF-print dendrogram tree. Results VNTR dendrogram of clustered VNTR genotypes indicated the two strains of Brucella isolated from Shanghai belonged to Brucella suis biover 2. Conclusions HOOP-Prints analysis is promising for investigating the molecular epidemiological profiles of Brucella infection in China.