布鲁氏菌分离株MLVA分子分型鉴定

Identification and classification of Brucella strain with MLVA

目的对牛奶、羊奶以及羊流产胎儿中分离布鲁氏菌进行分型鉴定。方法采集新疆4个地区牛奶20份,羊奶20份,羊流产胎儿10份,分离培养后,运用VirB8-PCR和布鲁氏菌分子分型PCR(AMOS-PCR)方法对分离株进行布鲁氏菌属、种的鉴定,同时采用MLVA方法对分离株进一步鉴定,并将测定结果与http://mlva.u-psud.fr/数据库提供的布鲁氏菌数据进行比较,结合软件BioNumerics 6.6进行聚类分析。结果 6株分离株均为羊种3型布鲁氏菌,与辽宁省分离羊种3型布鲁氏菌在聚类分析中关系较近。结论 MLVA作为布鲁氏菌基因分型鉴定的一种快速、可靠的方法,为今后布鲁氏菌传染源的追溯及防控措施的制定提供依据。

We identified the type of Brucellastrain from milk and abortus.The 20 portions of milk from dairy cows,20 portions of milk and 10 aborted fetuses from sheep were collected from four different area in Xinjiang.The suspected strains were identified by VirB8-PCR and AMOS-PCR.The further comparative analysis was conducted between our results and Brucella from database.（http：//mlva.u-psud.fr/）,and further phylogenetic tree was constructed by BioNumerics 6.6.Results showed that six Brucellastrain were identified as Brucella melitensis 3by MLVA,and were close to the Brucellastrain belonging to Liaoning Province.Multiple Loci VNTR Analysis（MLVA）assays could be applied fast and reliably to detect Brucella.The result of MLVA provide the basis for tracing the source of infection and the scientific basis for the establishment of comprehensive measures for prevention and control of brucellosis.