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目的对广东省羊种布鲁氏菌进行多位点序列分型(MLST)研究,了解菌株的遗传特征及遗传进化关系。方法采用MLST技术对60株羊种布鲁氏菌地方株的7个看家基因、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标准菌株的等位基因型进行比对,确定其等位基因谱型及菌株序列型(STS),分析与其他ST型间的遗传关系。结果 60株地方株经PCR产物检测、测序及序列分析,全部菌株的各等位基因谱相同,均为gap(3)、aro A(2)、glk(3)、dna K(2)、gyr B(1)、trp E(5)、cob Q(3)、omp25(8)、Int-hyp(2),与文献报道的ST8型的各等位基因型一致。经MLST技术分析聚类后,60株羊种布氏菌地方株与16株各生物种型标准株间的相似值介于2.5%~100.0%,100.0%相似值水平的菌株分成12种ST型别,其中60株地方株与羊种2型标准株同为ST8型。结论广东省羊种布鲁氏菌MLST基因分型的优势基因型别为ST8型,MLST分型技术不能有效区分广东省羊种布鲁氏菌的各生物型,只能作为羊种各生物型菌株分子分型的补充手段。
Objective To investigate the multi-locus sequence typing (MLST) of Brucella brucei in Guangdong Province and to understand the genetic characteristics and genetic evolution of the strains. Methods The sequences of seven housekeeping genes, one outer membrane protein gene and one intergenic region of 60 strains of brucella local strains were determined by MLST technique. The sequences of each gene were compared with those of the standard strains Genotypes were compared to determine the allelic genotype and strain sequence type (STS), analysis and other ST-type genetic relationship between. Results Sixty isolates of local strains were detected by PCR products, sequenced and sequenced. The alleles of all the isolates were the same (gap (3), aro A (2), glk (3), dna K B (1), trp E (5), cob Q (3), omp25 (8) and Int-hyp (2) were consistent with the reported allelic types of ST8. After clustering by MLST technique, the similarities between the 60 strains of brucella and 16 strains of each species were between 2.5% and 100.0%. The strains with 100.0% similarity level were divided into 12 ST types Do not, of which 60 strains of local strains and sheep species 2 standard strains with ST8 type. Conclusion The dominant genotype of Brucella MLST genotype in Guangdong Province is ST8 type. The MLST genotyping technique can not effectively distinguish the various genotypes of Brucella brucei in Guangdong Province, Strain molecular typing supplementary means.