基于经典单核苷酸多态性和多位点可变数目串联重复序列分析的中国炭疽芽胞杆菌基因遗传特征分析

Canonical single nucleotide polymorphism analysis and multiple-locus variable number tandem repeat analysis on genetic characteristics of Bacillus anthracis in China

  • 摘要:
    目的 分析中国炭疽芽胞杆菌基因型多态性,掌握中国炭疽芽胞杆菌基因遗传特征。
    方法 收集中国20个省份的炭疽芽胞杆菌菌株,采用经典单核苷酸多态性(canSNP)和多位点可变数目串联重复序列分析(MLVA)进行基因型鉴定,分析不同分离来源菌株基因型的分布特征;采用Bionumerics 5.1软件进行聚类分析,并计算各位点的Simpson指数。
    结果 533株中国炭疽芽胞杆菌均属于全球炭疽菌基因谱系中的A群,并具有全球12种亚群中的6种亚群,分别为A.Br.001/00 2、A.Br.Ames、A.Br.Aust94、A.Br.008/009、A.Br.Vollum和A.Br.005/006,其中A.Br.001/002亚群(60.98%)是中国的主要流行谱系,各地区均有分布,A.Br.Aust94(14.26%)、A.Br.008/009(11.82%)和A.Br.Vollum(4.69%)3个亚群主要分离自1980和1990年代的新疆维吾尔自治区(新疆),A.Br.Ames亚群菌株主要来自内蒙古自治区,主要分离自2010年代,A.Br.005/006亚群菌株最少,分离自2019年后。 canSNP亚群在中国的地理分布不均衡,新疆、广西壮族自治区、青海省、内蒙古自治区、河北省等具有4~5种亚群,而四川省等10省仅有1种亚群,新疆以A.Br.Aust94和A.Br.008/009为优势亚群,其他地区均以A.Br.001/002为优势亚群。 不同分离年代菌株的亚群分布有差异,在以A.Br.001/002为主要流行亚群的基础上,1980和1990年代A.Br.008/009、A.Br.Vollum和A.Br.Aust94相继出现并流行传播,2000年后A.Br.Ames亚群逐渐传播扩散,2019年A.Br.005/006亚群被检测到,显示了炭疽菌株基因型的传播和变异趋势。 通过MLVA15分型研究菌株具有91个基因型,结果与canSNP亚群基本一致,且各亚群内MLVA15的分型能力均较好。
    结论 本研究建立了中国炭疽芽胞杆菌canSNP和MLVA基因型数据库,掌握了中国炭疽杆菌基因型分布特征及其传播扩散趋势,为炭疽疫情防控及分子溯源技术提供了参考依据。

     

    Abstract:
    Objective To understand the genetic characteristics of Bacillus anthracis in China, and analyze the genotype polymorphism of B. anthracis.
    Methods A total of 533 isolates of B. anthracis were collected in China. Genotype identification was carried out by using the canonical single nucleotide polymorphism(canSNP)analysis and multiple-locus variable number tandem repeat analysis (MLVA) on genotype polymorphism genotyping protocol based on 15 variable number tandem repeat (VNTR) loci. Software Bionumerics 5.1was used for clustering analysis and minimum spanning trees were constructed to analyze the genetic relationships and Simpson’s index of each loci was calculated.
    Results All the 533 strains of B. anthracis belonged to the linage A of the global anthrax lineages, and had 6 sublineages, which were A.Br.001/002, A. Br.Ames, A.Br.008/009, A. Br.Vollum, A. Br.Aust94 and A. Br.005/006, respectively. Of these sublineages, A.Br.001/002 (60.98%) was predominant and widely distributed in China. A. Br.Aust94 (14.26%), A.Br.008/009 (11.82%) and A.Br.Vollum (4.69%) were mainly isolated from Xinjiang Uygur Autonomous Region (Xinjiang) in 1980s and 1990s, and A.Br.Ames was mainly isolated from Inner Mongolia. A.Br.005/006 was a newly discovered sublineage in 2019. The geographical distribution of sublineages of B. anthracis varied with province, with 4 - 5 sublineages in Xinjiang, Guangxi Zhuang Autonomous Region, Qinghai, Inner Mongolia Autonomous Region and Hebei, and only 1 sublineage in other 10 provinces. The predominant sublineages of B. anthracis were A.B.Aust94 and A.B.008/009 in Xinjiang, and A.B.001/002 in other regions. The distribution of sublineages were different among years. A. Br.001/002 was the predominant sublineage, and A. Br.008/009, A. Br.vollum and A. Br.Aust94 emerged and spread successively in the 1980s and 1990s, A .Br. Ames gradually spread after 2000, and a new sublineage A.Br.005/006 was detected in 2019. The study result revealed the genetic characteristics and the spread trend of B. anthracis in China. There were 91 genotypes indicated by MLVA15, and the results of MLVA15 genotyping were basically consistent with canSNP typing. The high resolution ability of MLVA15 was found in all sublineages.
    Conclusion In this study, the canSNP and MLVA genotype databases of B. anthracis were established in China, and the distribution characteristics and spread trend of B. anthracis were revealed, which provided scientific basis for the control and molecular source tracing of the anthrax outbreak.

     

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