Development of a Sandwich ELISA for EHEC O157:H7 Intimin gamma 1

文献类型： 外文期刊

第一作者： Zhang, Xuehan

作者： Zhang, Xuehan;Li, Meng;Zhang, Bicheng;He, Kongwang;Zhang, Xuehan;Li, Meng;Zhang, Bicheng;He, Kongwang;Zhang, Xuehan;Li, Meng;Zhang, Bicheng;He, Kongwang;Chen, Kangming

作者机构：

期刊名称：PLOS ONE （ 影响因子：3.24； 五年影响因子：3.788 ）

ISSN： 1932-6203

年卷期： 2016 年 11 卷 9 期

页码：

收录情况： SCI

摘要： Enterohemorrhagic Escherichia coli (EHEC) O157:H7 is a zoonotic pathogen of worldwide importance that causes foodborne infections in humans. Intimin gamma 1 (intimin gamma 1) is one of the most important outer membrane proteins required for EHEC's intimate adhesion to epithelial cells. Here, we generated a polyclonal antibody (pAb) and a monoclonal antibody (mAb) against intimin gamma 1 to develop a double antibody sandwich ELISA (DAS-ELISA) with increased sensitivity and specificity for measuring EHEC O157: H7. To achieve this goal, a rabbit pAb was used as a capture antibody, and a mouse mAb was a detection antibody. No cross-reactivity was observed with the other genera of pathogenic bacteria tested with the DAS-ELISA, which included Salmonella enteritidis, Shigella flexneri type 2, Listeria monocytogenes, Streptococcus suis type 2, and other 18 serotype E. coli. Detection limits of the DAS-ELISA were 1 x 10(3) CFU/mL for EHEC O157:H7 cultures, 1 x 10(4) CFU/g before enrichment, and 1 x 10(2) CFU/g after enrichment of contaminated samples. Field samples (n = 498) were tested using a previously established duplex-PCR method and compared to our DAS-ELISA. The DAS-ELISA had a specificity of 94.4%, a sensitivity of 91.5% and accuracy of 94.0% compared with duplex-PCR. The DAS-ELISA developed here can be applied to EHEC O157:H7 quantification in food, animal, and environmental samples.

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