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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。
Objective To understand the sequence type (ST) and genetic characteristics of Brucella isolates from goats isolated in Guizhou Province in 2010, and provide a scientific basis for the prevention and control of brucellosis among animals and humans. Methods Two strains of Brucella isolated from goats in 2010 were identified by using Brucella-specific PCR (BCSP31-PCR) and species-specific PCR (AMOS-PCR) locus sequence typing (MLST) technique was used to determine the sequence of seven housekeeping genes, one outer membrane protein gene and one intergenic region of two strains of Brucella. The sequence of each gene was compared with the allele of the reference strain The alleles and sequence types (STs) of these strains were determined. The genetic evolution relationship between ST strains of two strains and representative strains of Brucella strains was analyzed. Results Two Brucella strains from goats in Guizhou Province were identified as Brucella by BCSP31-PCR, and Brucella was identified by AMOS-PCR. MLST analysis revealed that the nine alleles of the two strains were of the ST8 type and most closely related to Brucella ST7, ST9-ST12, ST34 and ST35 of the same species. Conclusion The two Brucella isolates isolated in 2010 in Guizhou Province are all Brucella ST8 strains. The genetic relationship with the ST genotype of the same species of Brucella is the most recent, and the result is the prevention and control of brucellosis among humans and animals in Guizhou Province Control provides a scientific basis.