食品科学

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2008—2012年上海市肠炎沙门氏菌分离株毒力基因筛查与ERIC-PCR分型

庄孝飞1,周秀娟1,许学斌2,史贤明1,施春雷1,*   

  1. 1.上海交通大学农业与生物学院,中美食品安全联合研究中心,陆伯勋食品安全研究中心,
    微生物代谢国家重点实验室,上海 200240;2.上海市疾病预防控制中心,上海 200336
  • 出版日期:2015-07-25 发布日期:2015-07-15
  • 通讯作者: 施春雷
  • 基金资助:

    国家高技术研究发展计划(863计划)项目(2012AA101601);“十二五”国家科技支撑计划项目(2012BAK17B10);
    上海市国际合作项目(14390711900)

Screening of Virulence Genes and ERIC-PCR Sub-typing forS almonella Enteritidis Isolates in Shanghai during 2008—2012

ZHUANG Xiaofei1, ZHOU Xiujuan1, XU Xuebin2, SHI Xianming1, SHI Chunlei1,*   

  1. 1. MOST-USDA Joint Research Center for Food Safety, Bor Luh Food Safety Center, State Key Laboratory of Microbial Metabolism,
    School of Agriculture and Biology, Shanghai Jiao Tong University, Shanghai 200240, China;
    2. Center for Disease Control and Prevention of Shanghai, Shanghai 200336, China
  • Online:2015-07-25 Published:2015-07-15
  • Contact: SHI Chunlei

摘要:

采集上海地区2008—2012年来自临床病人和市售食品的肠炎沙门氏菌分离株90 株;采用聚合酶链式反应(polymerase chain reaction,PCR)方法,对存在于毒力岛SPI和毒力质粒上的9 种常见毒力基因携带情况进行了调查;并采用本实验优化的肠杆菌基因间保守重复序列-聚合酶链式反应(enterobacterial repetitive intergenicconsensus-PCR,ERIC-PCR)方法对这些分离株进行亚分型。结果显示:毒力基因invH、sopE、sugR的携带率为100.0%(90/90),iacP、avrA、rhuM、prgK均为98.9%(89/90),而spvB、spvC分别为85.6%(77/90)和78.9%(71/90)。优化的ERIC-PCR体系能够较好地区分8 种主要沙门氏菌血清型,共17 株菌,辛普森指数(D值)为0.970 6。然而,90 株肠炎沙门氏菌分离株却具有一致的ERIC-PCR指纹图谱。在所调查分离株中,毒力基因的携带率较高,尤其是invH、sopE和sugR,对人类健康存在较大威胁;ERIC-PCR虽然可用于区分沙门氏菌不同血清型,但对肠炎沙门氏菌的分型效果不佳。

关键词: 肠炎沙门氏菌, 毒力基因, ERIC-PCR分型

Abstract:

In this study, a total of 90 isolates of Salmonella Enteritidis, including clinical and foodborne isolates, were
collected from different areas of Shanghai during 2008 to 2012. Polymerase chain reaction (PCR) was used to investigate the
carrying rates of 9 common virulence genes not only in SPIs (SPI-1 and SPI-3) but also in virulence plasmid. In addition, an
optimized enterobacterial repetitive intergenic consensus (ERIC)-PCR was applied to subtype these isolates. The PCR results
of 90 isolates showed that the carrying rate was 100.0% for virulence genes invH, sopE and sugR, and 98.9% for iacP, avrA,
prgK and rhuM, while the carrying rate of spvB and spvC was 85.6% and 78.9% respectively. Seventeen strains belonging
to eight different Salmonella serovars were clearly differentiated by ERIC-PCR, with the Simpson’s Diversity Index up to
0.970 6. However, the ERIC-PCR profiles of 90 Salmonella Enteritidis isolates showed no difference. In conclusion, the
carrying of these virulence genes, especially sugR, invH and sopE, is very common among these isolates, demonstrating that
they pose a potential threat to human health and that ERIC-PCR can be applied to the molecular classification of different
Salmonella serovars, but not for Salmonella Enteritidis.

Key words: Salmonella Enteritidis, virulence genes, ERIC-PCR

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