沙门氏菌属特异性检测靶点碱基差异位点的识别及其血清型决定力的分析

食品科学 ›› 0, Vol. ›› Issue (): 0-0.

• 生物工程 • 下一篇

沙门氏菌属特异性检测靶点碱基差异位点的识别及其血清型决定力的分析

石秀清¹,史贤明²,周秀娟²,施春雷³

1. 上海交通大学
2. 上海交通大学农业与生物学院
3. 上海交通大学农业与生物学院食品科学系

收稿日期:2016-12-20 修回日期:2017-09-25 出版日期:2017-12-25 发布日期:2017-12-07
通讯作者: 石秀清 E-mail:shixiuqing@sjtu.edu.cn
基金资助:
政府间国际科技创新合作重点专项

Identification of SNPs in Different Serotypes of Salmonella-specific Targets and Its Analysis for The Determinationof Molecular Serotyping

Received:2016-12-20 Revised:2017-09-25 Online:2017-12-25 Published:2017-12-07
Contact: Xiu-qing SHI E-mail:shixiuqing@sjtu.edu.cn
Supported by:
Program of international S&T cooperation

摘要/Abstract

摘要： 沙门氏菌是引发食物中毒最主要的致病菌之一，复杂多样的血清型增加了对它检测和控制的难度，开发快速准确的沙门氏菌检测与分子血清分型方法对沙门氏菌的防控具有重要意义。本文以NCBI上已公布的28株沙门氏菌（14个血清型）全基因组序列为研究对象，对前期研究获得的7个沙门氏菌属特异性检测靶点在不同血清型间的单核苷酸多态性（SNPs）进行了比较分析，结果表明，S9和S69为碱基差异位点最多的两个靶点，具有沙门氏菌分子血清分型的潜在能力。随后，通过分析S9和S69在沙门氏菌不同血清型菌株中的差异位点，分别绘制两个血清分型靶点的差异位点表，从而获得了这两个靶点同时用于14个沙门氏菌血清型的快速分子血清分型的差异位点组合。在这些组合中，同一血清型具有相同的差异位点，不同血清型可以通过差异位点得以区分。最后，采集食品样品192份，用国标方法获得疑似沙门氏菌21株；利用血清分子分型靶点S9和S69进行快速分型，并同时用传统玻片凝集反应鉴定方法进行验证，两种方法鉴定结果的符合率为100%。鉴定结果为：21株沙门氏菌共包括4个不同血清型，其中肠炎沙门氏菌10株，鼠伤寒沙门氏菌7株，猪霍乱沙门氏菌2株，纽波特沙门氏菌2株。基因组序列分析结果和分离株分型结果均表明，沙门氏菌特异分子检测靶点S9和S69相结合具备沙门氏菌的分子血清分型能力，以差异位点表为基础建立的血清分型方法有望替代传统的玻片凝集血清分型这一繁杂步骤，从而降低沙门氏菌血清鉴定的时间与成本，为PCR技术应用于沙门氏菌分子血清分型提供新思路。

关键词: 沙门氏菌, 分子血清分型, 特异性检测靶点, 食品样品, 血清鉴定

Abstract: Salmonella is one of the main pathogenic bacteria causing food poisoning. These complex serotypes are important indicators for its detection. Development of fast and accurate Salmonella detection and serotyping methods are important for the prevention and control of Salmonella outbreak. The single nucleotide polymorphism (SNPs) in different serotypes of 7 Salmonella-specific detection targets were analyzed based on the whole genome sequence of 28 Salmonella strains (14 serotypes) downloaded from NCBI. The results showed that S9 and S69 have more SNPs than others, which have the potentiality for the Salmonella serotyping. Subsequently, nucleotide base site difference table of the two candidate molecular serotyping targets were drawn by sorting out the differences of S9 and S69 in the Salmonella strains of different serotypes strains, respectively. Differential site combination of two target points for rapid molecular serotyping of 14 Salmonella serotypes, and shows that identical mutation sites existed in the same serovar, while different mutation sites were found in diverse serovars. Finally, 192 food samples were collected from supermarkets and open-air markets in Shagnhai, and 21 suspected Salmonella isolates were obtained. The results of molecular serotyping by targets S9 and S69 were consistent with traditional slide agglutination tests (coincidence rate 100%), including 4 different serovars:10 strains of Salmonella Enteritidis, 7 strains of Salmonella Typhimurium, 2 strains of Salmonella Choleraesuis and 2 strains of Salmonella Newport. The results of genome sequence analysis and strain serotyping showed that combination of Salmonella-specific molecular detection targets S9 and S69 has the molecular serotyping capacity for Salmonella, which is expected to be an alternative method for traditional of slide agglutination. It will reduce the time and cost based on the nucleotide base site difference table, which alsoprovides new ideas for the application of PCR technology in Salmonella molecular serotyping.

Key words: Salmonella, Molecular Serotyping, Salmonella-specific detection Targets, Food Samples, Serum Identification

中图分类号:

Q781

石秀清史贤明周秀娟施春雷. 沙门氏菌属特异性检测靶点碱基差异位点的识别及其血清型决定力的分析[J]. 食品科学, 0, (): 0-0.

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沙门氏菌属特异性检测靶点碱基差异位点的识别及其血清型决定力的分析

Identification of SNPs in Different Serotypes of Salmonella-specific Targets and Its Analysis for The Determinationof Molecular Serotyping

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