Multilocus sequence typing method for identification and genotypic classification of pathogenic Leptospira species.
Multilocus sequence typing method for identification and genotypic classification of pathogenic Leptospira species.
复制标题
DOI：
10.1186/1476-0711-5-28
复制
发表时间：
2006-11-23
影响因子：
5.7
通讯作者：
Hartskeerl, Rudy A
中科院分区：
文献类型：
--
作者：
Ahmed, Niyaz;Devi, S Manjulata;Hartskeerl, Rudy A
背景：钩端螺旋体是与广泛的哺乳动物宿主相关的寄生细菌有机体，是导致人类钩端螺旋体病严重病例的原因。钩端螺旋体病的流行病学是复杂和动态的。已经确定了多种血清型，每种血清型都适应于一种或多种动物宿主。适应是一个动态的过程，它改变了血清型和临床表现在不同宿主体内的时空分布。基于表面抗原库的血清分型是钩端螺旋体病原体分类的模糊和人工系统。病原钩端螺旋体的分子分型方法自全基因组序列的破译以来一直受到高度关注。目前，基于多位点序列分型（MLST）等单独技术的微生物基因型数据资源很少，但不幸的是，没有这样的钩端螺旋体数据库。结果：我们首次报道了钩端螺旋体基因分型的可靠MLST方法的发展。采用4个管家基因和2个候选基因的DNA序列一致性进行分型分析，其中包括来自不同地理区域和不同来源的41个参考菌株。在6个选择的基因中，adk、icdA和secY的变异性较大，而LipL32和LipL41编码基因以及rrs2基因的变异性较小。系统发育树根据基于基因组的物种对分离物进行聚类。结论：MLST与其他钩端螺旋体分型方法相比，具有重现性、稳健性、一致性和可移植性等优点。钩端螺旋体的遗传亲缘关系可以通过MLST方法更好地研究，并可用于分子流行病学和进化研究以及群体遗传学。
BACKGROUND: Leptospira are the parasitic bacterial organisms associated with a broad range of mammalian hosts and are responsible for severe cases of human Leptospirosis. The epidemiology of leptospirosis is complex and dynamic. Multiple serovars have been identified, each adapted to one or more animal hosts. Adaptation is a dynamic process that changes the spatial and temporal distribution of serovars and clinical manifestations in different hosts. Serotyping based on repertoire of surface antigens is an ambiguous and artificial system of classification of leptospiral agents. Molecular typing methods for the identification of pathogenic leptospires up to individual genome species level have been highly sought after since the decipherment of whole genome sequences. Only a few resources exist for microbial genotypic data based on individual techniques such as Multiple Locus Sequence Typing (MLST), but unfortunately no such databases are existent for leptospires.RESULTS: We for the first time report development of a robust MLST method for genotyping of Leptospira. Genotyping based on DNA sequence identity of 4 housekeeping genes and 2 candidate genes was analyzed in a set of 120 strains including 41 reference strains representing different geographical areas and from different sources. Of the six selected genes, adk, icdA and secY were significantly more variable whereas the LipL32 and LipL41 coding genes and the rrs2 gene were moderately variable. The phylogenetic tree clustered the isolates according to the genome-based species.CONCLUSION: The main advantages of MLST over other typing methods for leptospires include reproducibility, robustness, consistency and portability. The genetic relatedness of the leptospires can be better studied by the MLST approach and can be used for molecular epidemiological and evolutionary studies and population genetics.