贵州省两株山羊源羊种布鲁菌MLST分析

目的了解贵州省2010年分离自山羊的两株羊种布鲁菌的等位基因序列型(Sequence type, ST)及遗传学特征,为动物间和人间布病疫情的预防和控制提供科学依据。方法应用布鲁菌属特异性PCR(BCSP31-PCR)和种/型特异性PCR(AMOS-PCR)对2010年分离自山羊的两株布鲁菌进行鉴定,采用多位点序列分型(Multiple locus sequence typing, MLST)技术对两株布鲁菌菌株的7个管家基因、1个外膜蛋白基因及1个基因间区的序列进行测定,将各个基因的序列与参考菌株的等位基因型进行比对,确定其等位基因谱及序列型(STs),分析与两株菌ST型与各种型布鲁菌代表菌株的遗传进化关系。结果来自贵州省山羊的两株布鲁菌株经BCSP31-PCR鉴定为布鲁菌属细菌,AMOS-PCR将其鉴定为羊种布鲁菌。MLST分析显示,两株菌的9个等位基因序列型为ST8型,与同属羊种布鲁菌的ST7、ST9~ST12、ST34和ST35遗传关系最近。结论贵州省2010年分离的2株布鲁菌分离株均为ST8型布鲁菌,与同属羊种布鲁菌的ST型遗传关系最近,结果为贵州省人间和动物间布病疫情的预防和控制提供了科学依据。

Multiple locus sequence typing of two strains of Brucella melitensis isolated from goats in Guizhou Province

In order to understand the allele locus gene sequence type (ST) and the genetic characteristic of two Brucella melitensis strains isolated from goats in Guizhou Province in 2010 and provide scientific basis for the control and prevention of animal and human brucellosis, the two isolates were identified by using Brucella genus specific BCSP31-PCR and species-specific AMOS-PCR, and the seven house-keeping genes and one out membrane protein gene and one gene -inter- region sequence of the strains were further analyzed using multiple locus sequence typing (MLST). Each gene sequence was compared with the reference strains to determine the allelic profile and sequence type, and the ST data-based genetic relationship of the two isolates and Brucella strains representing different species were analyzed. The results showed that the two Brucella strains were identified as B. melitensis with BCSP31-PCR and AMOS-PCR. MLST analysis revealed that the 9 alleles based sequence type of the two strains was ST8, which was genetically closed to the STs (ST7, ST8-ST12, ST34 and ST35) belonging to B. melitensis. The results indicate that the sequence type of the two strains of B. melitensis isolated from goats in Guizhou Province in 2010 is ST8, close to the STs belonging to B. melitensis.