| 摘要: |
| 【目的】建立一种快速检测动物源大肠埃希菌O157∶H7及其毒力基因的多重PCR方法。【方法】以编码大肠埃希菌O157抗原的rfbE基因、编码H7抗原的fliC基因以及编码细胞溶血素A的hylA基因为靶基因，设计3对特异性引物，优化并建立检测大肠埃希菌O157∶H7的多重PCR方法，对其特异性和敏感性进行检测，最后对其临床应用效果进行了初步验证。【结果】成功建立了快速检测和鉴定动物源大肠埃希菌O157∶H7 rfbE、fliC和hylA基因的多重PCR方法，该方法特异性较好，灵敏度较高，可达2.0×102 CFU/mL。【结论】初步建立了检测动物源大肠埃希菌O157∶H7的多重PCR方法，该方法可用于携带大肠埃希菌O157∶H7临床动物的分子流行病学调查。 |
| 关键词: 大肠埃希菌O157∶H7 多重PCR rfbE基因 fliC基因 hylA基因 |
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| 基金项目:河南省重大公益科研项目(81100912300)；国家科技支撑计划项目(2007BAQ01047)；漯河市科技计划项目(081203) |
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| Preliminary study on multiplex PCR for detecting animal derived Escherichia coli O157∶H7 |
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HU Hui1, DUAN Zhigang1, CUI Baoan,et al2
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1.College of Animal Science and Veterinary Medicine ,Henan Argricultural University;2.College of Animal Science and Veterinary Medicine, Henan Argricultural University
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| Abstract: |
| 【Objective】The multiplex PCR for detecting Escherichia coli O157∶H7 antigens and virulence gene hylA was established.【Method】3 pairs of specific primers were designed according to the sequence of antigen genes of rfbE,fliC and virulence gene hylA of E.coli O157∶H7.And the multiple PCR was constructed using these primers in one amplificated system by optimizing the amplificated conditions.The specificity and sensitivity of this system were evaluated,and used to detect the clinical isolates.【Result】The multiplex PCR method was established successfully,which could simultaneously detect the O-antigen,H-antigen and virulence gene hylA of E.coli O157∶H7 with a high specificity and sensitivity.Sensitivity of the assay was 2×102 CFU/mL of bacteria samples.【Conclusion】The established multiplex PCR method could be used in clinical diagnosis and epidemiological investigation of animal-derived E.coli O157∶H7. |
| Key words: Escherichia coli O157∶H7 multiplex PCR rfbE gene fliC gene hylA gene |
