| 多重PCR-DHPLC技术检测肠出血性大肠杆菌O157:H7基因型的方法学研究 |
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| 引用本文： | 徐君怡,曹际娟,郑秋月,王秋艳,刘淑艳,蒋丹.多重PCR-DHPLC技术检测肠出血性大肠杆菌O157:H7基因型的方法学研究[J].中华微生物学和免疫学杂志,2009,29(4). |
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| 作者姓名： | 徐君怡 曹际娟 郑秋月 王秋艳 刘淑艳 蒋丹 |
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| 作者单位： | 辽宁出入境检验检疫局技术中心食品微生物与转基因检测国家重点实验室,大连,116015 |
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| 摘 要： | 目的 开发肠出血性大肠杆菌O157:H7的多重PCR-变性高效液相色谱(DHPLC)检测方法 .方法 以编码大肠杆菌0157抗原的rfbE 基因、编码毒力因子的类志贺毒素(SLT)基因为目的 基因,选择2对引物,建立并优化了大肠杆菌O157:H7的多重PCR-DHPLC检测体系.扩增产物分别为224 bp和499 bp.结果 采用37株细菌验汪了该多重PCR具有良好的特异件.PCR榆测的灵敏度可达到4 CFU/ml.结论 实验证明,研究建立的多重PCR-DHPLC方法 可特异、灵敏地实现对大肠杆菌O157:H7的检测.
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| 关 键 词： | 大肠杆菌O157:H7 多重PCR 变性高效液相色谱 |
Detection of Escherichia coli O157:H7 by multiplex PCR-DHPLC |
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| XU Jun-yi,CAO Ji-juan,ZHENG Qiu-yue,WANG Qiu-yan,LIU Shu-yan,JIANG Dan.Detection of Escherichia coli O157:H7 by multiplex PCR-DHPLC[J].Chinese Journal of Microbiology and Immunology,2009,29(4). |
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| Authors: | XU Jun-yi CAO Ji-juan ZHENG Qiu-yue WANG Qiu-yan LIU Shu-yan JIANG Dan |
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| Abstract: | Objective To develop the PCR-denaturing high performance liquid chromatography (DHPLC) for detection of E. Coli O157: H7. Methods The virulence genes of Shiga-like toxin(SLT) and rfbE were specifically amplified by 2 sets of primers. The target gene fragments of the PCR assay were 224 bp and 499 bp, respectively. Results Analysis of 37 strains demonstrated that this PCR system was specific. The detection limit of the PCR was 4 CFU/ml. Conclusion These results indicated that the multiplex PCR-DHPLC assay can be used for specific and sensitive detection of E. Coli O157:H7. |
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| Keywords: | E coli O157: H7 Multiplex PCR Denaturing high performance liquid chromatogra-phy |
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