Characterization of Genetic Diversity of Bacillus anthracis in France by Using High-Resolution Melting Assays and Multilocus Variable-Number Tandem-Repeat Analysis
Characterization of Genetic Diversity of Bacillus anthracis in France by Using High-Resolution Melting Assays and Multilocus Variable-Number Tandem-Repeat Analysis
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DOI：
10.1128/jcm.05439-11
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发表时间：
2011-12-01
影响因子：
9.4
通讯作者：
Madani, N.
中科院分区：
文献类型：
--
作者：
Derzelle, S.;Laroche, S.;Madani, N.
利用高分辨熔融(HRM)分析，我们开发了一种经济高效的方法来对炭疽杆菌基因组中的13个具有系统发育信息的单核苷酸多态(SNPs)进行基因分型。对1953-2009年间在法国采集的100株炭疽杆菌和几个参考菌株进行了单、双链SNP鉴别分析。人力资源管理提供了一种可靠和廉价的替代炭疽亚型进入12个主要亚系或亚组之一。除巴斯德疫苗株ATCC 4229外，所有毒株均能正确定位在典型SNP(CanSNP)系统发育树上。我们在法国检测到了三个canSNP亚群的共存。在阿尔卑斯山、比利牛斯山、奥弗涅地区和Saone-et-Loire省发现了占优势的B.Br.CNEVA亚系。在全国大部分地区都观察到了属于A.Br.008/009亚组的菌株。较小的A.Br.001/002亚群仅限于法国东北部。使用24个标记的多位点可变数目串联重复分析进一步将法国菌株分解为60个独特的图谱，并确定了一些区域模式。在A.BR中发现了多样性。008/009和B.Br.CNEVA亚群表明，这些亚群代表着法国在生态上建立起来的古老支系。文中还讨论了与世界其他地区菌株的亲缘关系。
Using high-resolution melting (HRM) analysis, we developed a cost-effective method to genotype a set of 13 phylogenetically informative single-nucleotide polymorphisms (SNPs) within the genome of Bacillus anthracis. SNP discrimination assays were performed in monoplex or duplex and applied to 100 B. anthracis isolates collected in France from 1953 to 2009 and a few reference strains. HRM provided a reliable and cheap alternative to subtype B. anthracis into one of the 12 major sublineages or subgroups. All strains could be correctly positioned on the canonical SNP (canSNP) phylogenetic tree, except the divergent Pasteur vaccine strain ATCC 4229. We detected the cooccurrence of three canSNP subgroups in France. The dominant B.Br.CNEVA sublineage was found to be prevalent in the Alps, the Pyrenees, the Auvergne region, and the Saone-et-Loire department. Strains affiliated with the A.Br.008/009 subgroup were observed throughout most of the country. The minor A.Br.001/002 subgroup was restricted to northeastern France. Multiple-locus variable-number tandem-repeat analysis using 24 markers further resolved French strains into 60 unique profiles and identified some regional patterns. Diversity found within the A. Br. 008/009 and B. Br. CNEVA subgroups suggests that these represent old, ecologically established clades in France. Phylogenetic relationships with strains from other parts of the world are discussed.