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首页> 外文期刊>Journal of Clinical Microbiology >Rapid Multiplex PCR and Real-Time TaqMan PCR Assays for Detection of Salmonella enterica and the Highly Virulent Serovars Choleraesuis and Paratyphi C
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Rapid Multiplex PCR and Real-Time TaqMan PCR Assays for Detection of Salmonella enterica and the Highly Virulent Serovars Choleraesuis and Paratyphi C

机译：快速多重PCR和实时TaqMan PCR检测沙门氏菌和高毒血清型霍乱弧菌和副伤寒C

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摘要

Salmonella enterica is a human pathogen with over 2,500 serovars characterized. S. enterica serovars Choleraesuis and Paratyphi C are two globally distributed serovars. We have developed a rapid molecular-typing method to detect serovars Choleraesuis and Paratyphi C in food samples by using a comparative-genomics approach to identify regions unique to each serovar from the sequenced genomes. A Salmonella-specific primer pair based on oriC was designed as an internal control to establish accuracy, sensitivity, and reproducibility. Serovar-specific primer sets based on regions of difference between serovars Choleraesuis and Paratyphi C were designed for real-time PCR assays. Three primer sets were used to screen a collection of over 100 Salmonella strains, and both serovars Choleraesuis and Paratyphi C gave unique amplification patterns. To develop the technique for practical use, its sensitivity for detection of Salmonella spp. in a food matrix was determined by spiking experiments. The technique was also adapted for a real-time PCR rapid-detection assay for both serovars Choleraesuis and Paratyphi C that complements the current procedures for Salmonella sp. isolation and serotyping.
机译：沙门氏菌是一种人类病原体，具有2500多种血清型。 S。肠道病毒霍乱沙门氏菌和副伤寒C是两个全球分布的血清型。我们已经开发出一种快速的分子分型方法，通过使用比较基因组学方法从测序的基因组中鉴定出每个血清型所特有的区域，从而检测食品样品中的血清型霍乱弧菌和副伤寒C。设计基于 oriC 的沙门氏菌特异性引物对作为内部对照，以建立准确性，敏感性和可重复性。设计基于血清型霍乱弧菌和副伤寒C区差异区域的血清型特异性引物，用于实时PCR分析。使用三种引物组筛选了超过100种 Salmonella 菌株，并且血清型霍乱弧菌和副伤寒C都有独特的扩增模式。为了开发实用的技术，该技术可检测沙门氏菌（Salmonella） spp。通过加标实验确定食物基质中的维生素。该技术还适用于血清型霍乱沙门氏菌和副伤寒C的实时PCR快速检测测定法，以补充 Salmonella sp的当前程序。隔离和定型。

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