Use of a New High Resolution Melting Method for Genotyping Pathogenic Leptospira spp.
Use of a New High Resolution Melting Method for Genotyping Pathogenic Leptospira spp.
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DOI：
10.1371/journal.pone.0127430
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发表时间：
2015
期刊：
影响因子：
3.7
通讯作者：
Michault A
中科院分区：
文献类型：
--
作者：
Naze F;Desvars A;Picardeau M;Bourhy P;Michault A
钩端螺旋体病是一种世界性人畜共患病，在留尼汪岛等热带地区流行。问号钩端螺旋体（Leptospira interrogans）是人类感染的主要病原体，但其他病原体，如钩端螺旋体（Leptospira interrogans）是人类感染的主要病原体。kirschner和L. Borgpetersenii也与人类钩端螺旋体病有关。在这项研究中，用引物对lfb 1 F/R和G1/G2扩增产物的熔解曲线分析有助于钩端螺旋体参考菌株的准确种分类。接下来，我们结合了一种无监督的高分辨率熔解（HRM）方法与一种新的统计方法，使用引物扩增一个两个变量的串联重复序列（VNTR）在亚种水平上进行分型。使用ScreenClust软件进行的HRM分析使得能够以高分辨率（Hunter-Gaston指数0.984）在血清型水平鉴定基因型。这种方法也适用于1998年后从留尼汪岛获得的血液样本中的钩端螺旋体DNA。我们能够鉴定出一种独特的基因型，与L。问号血清型Copenhageni和黄疸出血型，表明这种基因型是留尼汪岛钩端螺旋体病的主要原因。我们的简单、快速和稳健的基因分型方法能够在种和亚种水平上鉴定钩端螺旋体菌株，并支持在不需要培养的情况下对生物样品中的钩端螺旋体进行直接基因分型。
Leptospirosis is a worldwide zoonosis that is endemic in tropical areas, such as Reunion Island. The species Leptospira interrogans is the primary agent in human infections, but other pathogenic species, such as L. kirschner and L. borgpetersenii, are also associated with human leptospirosis. In this study, a melting curve analysis of the products that were amplified with the primer pairs lfb1 F/R and G1/G2 facilitated an accurate species classification of Leptospira reference strains. Next, we combined an unsupervised high resolution melting (HRM) method with a new statistical approach using primers to amplify a two variable-number tandem-repeat (VNTR) for typing at the subspecies level. The HRM analysis, which was performed with ScreenClust Software, enabled the identification of genotypes at the serovar level with high resolution power (Hunter-Gaston index 0.984). This method was also applied to Leptospira DNA from blood samples that were obtained from Reunion Island after 1998. We were able to identify a unique genotype that is identical to that of the L. interrogans serovars Copenhageni and Icterohaemorrhagiae, suggesting that this genotype is the major cause of leptospirosis on Reunion Island. Our simple, rapid, and robust genotyping method enables the identification of Leptospira strains at the species and subspecies levels and supports the direct genotyping of Leptospira in biological samples without requiring cultures.
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影响因子：
2.2
作者：
Chan, Wai-Fong;Maharjan, Ram P.;Lan, Ruiting
通讯作者：
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实时PCR的开发和验证，用于检测临床材料中致病性钩端螺旋体。
影响因子：
3.7
作者：
Ahmed A;Engelberts MF;Boer KR;Ahmed N;Hartskeerl RA
通讯作者：
Hartskeerl RA
影响因子：
4.4
作者：
Desvars A;Michault A;Bourhy P
通讯作者：
Bourhy P
影响因子：
9.3
作者：
Gundry, CN;Vandersteen, JG;Wittwer, CT
通讯作者：
Wittwer, CT
影响因子：
9.4
作者：
HUNTER, PR;GASTON, MA
通讯作者：
GASTON, MA