华南预防医学 ›› 2016, Vol. 42 ›› Issue (5): 421-425.doi: 10.13217/j.scjpm.2016.0421

• 论著 • 上一篇    下一篇

广东省羊种布鲁氏菌的多位点序列分型研究

赖沛炼1,陈经雕2,彭晓放2,刘美真2,邓爱萍2,谢志坚3,黎薇2,管大伟2 ,肖红2 ,张万里2 ,苏娟2   

  1. 1.广州市中医医院,广东 广州 510010;2.广东省疾病预防控制中心;3.佛冈县疾病预防控制中心
  • 收稿日期:2016-03-31 出版日期:2016-11-16 发布日期:2016-12-02
  • 作者简介:赖沛炼( 1975―) ,男,大学本科,主管技师,主要从事临床病原微生物的实验室检测工作

Multilocus sequence typing of Brucella melitensis strains in Guangdong Province

LAI Pei-lian1,CHEN Jing-diao2,PENG Xiao-fang2,LIU Mei-zhen2,DENG Ai-ping2,XIE Zhi-jian3,LI Wei2,GUAN Da-wei2,XIAO Hong2,ZHANG Wan-li2,SU Juan2   

  1. Shenzhen Longgang District Center for Disease Control and Prevention,Shenzhen 518172, China
  • Received:2016-03-31 Online:2016-11-16 Published:2016-12-02
  • Contact: 广东省医学科研基金项目(No.A2014085) E-mail:1400137369@qq.com

摘要: 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)、aroA(2)、glk(3)、dnaK(2)、gyrB(1)、trpE(5)、cobQ(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。

Abstract: ObjectiveTo understand the hereditary character and evolutionary relationship of Brucella melitensis.MethodsThe multilocus sequence typing (MLST) assay was used to sequence 60 strains ofB. melitensis from Guangdong Province. Seven house keeping genes, 1 outer membrane gene, and 1 intergenic region of the 60 strains were sequenced and compared with alleles of standard strains to determine the sequence type. All the strains were analyzed through PCR detection, sequence and sequence analysis.ResultsThe sequences of all the 60 strains weregap(3), aroA(2), glk(3), dnaK(2), gyrB(1), trpE(5), cobQ(3), omp25(8), andInt-hyp(2), consistent with the various alleles of sequences type ST8 reported in the literature. The similarity among the 60 local strains and 16 standard ones ranged from 2.5% to 100.0%. The strains with 100.0% similarity were divided into 12 ST types, of which the 60 local strains were ST8 type, same asB. melitensis 2 standard strain. ConclusionB. melitensis ST8 is dominant in Guangdong. MLST is only a supplementary method but cannot be used to differentiateB. melitensis in Guangdong Province.

中图分类号: 

  • R446.5