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首页> 外文期刊>Journal of Clinical Microbiology >Development of a New Molecular Subtyping Tool for Salmonella enterica Serovar Enteritidis Based on Single Nucleotide Polymorphism Genotyping Using PCR
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Development of a New Molecular Subtyping Tool for Salmonella enterica Serovar Enteritidis Based on Single Nucleotide Polymorphism Genotyping Using PCR

机译：基于PCR的单核苷酸多态性基因分型的肠炎沙门氏菌肠炎沙门氏菌分子分型新工具的开发

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The lack of a sufficiently discriminatory molecular subtyping tool for Salmonella enterica serovar Enteritidis has hindered source attribution efforts and impeded regulatory actions required to disrupt its food-borne transmission. The underlying biological reason for the ineffectiveness of current molecular subtyping tools such as pulsed-field gel electrophoresis (PFGE) and phage typing appears to be related to the high degree of clonality of S. Enteritidis. By interrogating the organism's genome, we previously identified single nucleotide polymorphisms (SNP) distributed throughout the chromosome and have designed a highly discriminatory PCR-based SNP typing test based on 60 polymorphic loci. The application of the SNP-PCR method to DNA samples from S. Enteritidis strains (n = 55) obtained from a variety of sources has led to the differentiation and clustering of the S. Enteritidis isolates into 12 clades made up of 2 to 9 isolates per clade. Significantly, the SNP-PCR assay was able to further differentiate predominant PFGE types (e.g., XAI.0003) and phage types (e.g., phage type 8) into smaller subsets. The SNP-PCR subtyping test proved to be an accurate, precise, and quantitative tool for evaluating the relationships among the S. Enteritidis isolates tested in this study and should prove useful for clustering related S. Enteritidis isolates involved in outbreaks.
机译：缺乏针对肠炎沙门氏菌肠炎沙门氏菌的足够区分性的分子亚型分析工具，阻碍了源归因的努力，并阻碍了破坏其食源性传播所需的监管措施。当前分子亚分型工具（如脉冲场凝胶电泳（PFGE）和噬菌体分型）无效的潜在生物学原因似乎与 S 的高度克隆性有关。肠炎。通过询问生物体的基因组，我们先前确定了分布在整个染色体上的单核苷酸多态性（SNP），并基于60个多态性位点设计了基于高区分性PCR的SNP分型测试。 SNP-PCR方法在 S DNA样品中的应用。从各种来源获得的肠炎肠炎菌株（ n = 55）导致 S 的分化和聚集。肠炎肠球菌分为12个进化枝，每个进化枝2至9个分离菌组成。重要的是，SNP-PCR测定法能够将主要的PFGE类型（例如，XAI.0003）和噬菌体类型（例如，噬菌体类型8）进一步区分为较小的亚组。 SNP-PCR亚型测试被证明是评估 S 之间关系的准确，精确和定量的工具。肠炎沙门氏菌在本研究中进行了测试，应证明对与相关的 S 聚类有用。肠炎沙门氏菌与暴发有关。

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