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PCR-免疫胶体金试纸条方法检测食品中 肠出血性大肠杆菌O157:H7
引用本文:庞 璐,宋 喆,吴冬雪,徐宝东,张海英,赵良娟,刘 培,蔡国瑞,李宗梦,赵 宏,张宏伟. PCR-免疫胶体金试纸条方法检测食品中 肠出血性大肠杆菌O157:H7[J]. 食品安全质量检测学报, 2015, 6(2): 447-451
作者姓名:庞 璐  宋 喆  吴冬雪  徐宝东  张海英  赵良娟  刘 培  蔡国瑞  李宗梦  赵 宏  张宏伟
作者单位:天津出入境检验检疫局动植物与食品检测中心,天津出入境检验检疫局动植物与食品检测中心,天津出入境检验检疫局动植物与食品检测中心,天津出入境检验检疫局动植物与食品检测中心,天津出入境检验检疫局动植物与食品检测中心,天津出入境检验检疫局动植物与食品检测中心,天津出入境检验检疫局动植物与食品检测中心,天津出入境检验检疫局动植物与食品检测中心,天津出入境检验检疫局动植物与食品检测中心,天津出入境检验检疫局动植物与食品检测中心,天津出入境检验检疫局动植物与食品检测中心
基金项目:国家质检总局科技计划项目(2013IK181、2014IK222)
摘    要:目的建立PCR-免疫胶体金试纸条法快速检测食品中肠出血性大肠杆菌O157:H7的分析方法。方法通过设计特异性引物建立肠出血性大肠杆菌O157:H7 PCR检测方法并使用免疫胶体金技术以及双抗体夹心法建立PCR产物快速检测试纸条并设计核酸检测展开液;将1株大肠杆菌O157:H7标准菌株和7株其他常见食源性致病菌作为试验菌株,用试验菌株检测PCR-免疫胶体金试纸条方法的检测特异度,并比较PCR-免疫胶体金试纸条法和PCR-琼脂糖凝胶电泳法的检测敏感度。结果 PCR-免疫胶体金法具有良好的特异度,灵敏度比标准琼脂糖凝胶电泳法高100倍。结论本文建立的肠出血性大肠杆菌O157:H7检测PCR-免疫胶体金试纸条法特异度好,灵敏度高,价格低廉,适用于食品中肠出血性大肠杆菌O157:H7的检测。

关 键 词:肠出血性大肠杆菌O157:H7   快速检测   聚合酶链反应   免疫胶体金
收稿时间:2014-12-23
修稿时间:2015-02-06

Detection of Escherichia coli O157:H7 in food by PCR-immunogold method
PANG Lu,SONG Zhe,WU Dong-Xue,XU Bao-Dong,ZHANG Hai-Ying,ZHAO Liang-Juan,LIU Pei,CAI Guo-Rui,LI Zong-Meng,ZHAO Hong and ZHANG Hong-Wei. Detection of Escherichia coli O157:H7 in food by PCR-immunogold method[J]. Journal of Food Safety & Quality, 2015, 6(2): 447-451
Authors:PANG Lu  SONG Zhe  WU Dong-Xue  XU Bao-Dong  ZHANG Hai-Ying  ZHAO Liang-Juan  LIU Pei  CAI Guo-Rui  LI Zong-Meng  ZHAO Hong  ZHANG Hong-Wei
Affiliation:Animal, Plant and Foodstuffs Inspection Center, Tianjin Entry-Exit Inspection and Quarantine Bureau,Animal, Plant and Foodstuffs Inspection Center, Tianjin Entry-Exit Inspection and Quarantine Bureau,Animal, Plant and Foodstuffs Inspection Center, Tianjin Entry-Exit Inspection and Quarantine Bureau,Animal, Plant and Foodstuffs Inspection Center, Tianjin Entry-Exit Inspection and Quarantine Bureau,Animal, Plant and Foodstuffs Inspection Center, Tianjin Entry-Exit Inspection and Quarantine Bureau,Animal, Plant and Foodstuffs Inspection Center, Tianjin Entry-Exit Inspection and Quarantine Bureau,Animal, Plant and Foodstuffs Inspection Center, Tianjin Entry-Exit Inspection and Quarantine Bureau,Animal, Plant and Foodstuffs Inspection Center, Tianjin Entry-Exit Inspection and Quarantine Bureau,Animal, Plant and Foodstuffs Inspection Center, Tianjin Entry-Exit Inspection and Quarantine Bureau,Animal, Plant and Foodstuffs Inspection Center, Tianjin Entry-Exit Inspection and Quarantine Bureau and Animal, Plant and Foodstuffs Inspection Center, Tianjin Entry-Exit Inspection and Quarantine Bureau
Abstract:Objective To establish a PCR-immunogold strip method for rapid detection of Escherichia coli O157:H7 in food. Methods The PCR method was established through the design of specific primers and the establishment of rapid test immunogold strip for detection of PCR products combined immunogold skills with double antibody sandwich method. Test specificity of the new method using the test bacterial strains which include 1 Escherichia coli O157:H7 and 7 common food-borne pathogens and compare the sensitivities of the immunogold strip method and the traditional agarose gel electrophoresis method. Results The established method was specific and the sensitivity was higher than the agarose gel electrophoresis by 100 times. Conclusion The new method is suitable for the detection of Escherichia coli O157:H7 in food.
Keywords:Escherichia coli O157:H7   rapid detection   polymerase chain reaction   immunogold
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