| 环介导等温扩增技术检测小肠结肠炎耶尔森氏菌的研究 |
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| 引用本文： | 梁磊,李英军,孟兆祥,马晓燕,张伟.环介导等温扩增技术检测小肠结肠炎耶尔森氏菌的研究[J].食品科技,2011(9):335-339. |
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| 作者姓名： | 梁磊 李英军 孟兆祥 马晓燕 张伟 |
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| 作者单位： | 河北农业大学食品科技学院; |
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| 基金项目： | 河北省自然科学基金项目(C2008000216) |
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| 摘 要： | 采用环介导等温扩增技术(Loop-mediated isothermal amplification,LAMP)快速检测小肠结肠炎耶尔森氏菌(Yersinia enterocolitica)。以小肠结肠炎耶尔森氏菌(AY004311.1)Ail基因序列作为靶序列,设计内、外引物,通过肉眼观察沉淀判断检测结果。结果表明:LAMP检测小肠结肠炎耶尔森氏菌的灵敏度为6.3cfu/mL,人工污染鸡肉的检出限为340cfu/g。PCR检测小肠结肠炎耶尔森氏菌的灵敏度为630cfu/mL,人工污染鸡肉的检出限为3.4×104cfu/g。采用试剂盒法提取DNA,从样品处理到报告结果,LAMP方法耗时2h,PCR方法耗时3h。因此,LAMP检测小肠结肠炎耶尔森氏菌的灵敏度高,耗时短,特异性好,操作简便,无需特殊的仪器设备,适合在我国广大基层实验室开展应用,为快速检测食源性致病菌构建了一个新的技术平台。
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| 关 键 词： | 环介导等温扩增 检测 小肠结肠炎耶尔森氏菌 |
Development of a loop-mediated isothermal amplification forthe detection of Yersinia enterocolitica |
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| LIANG Lei,LI Ying-jun,MENG Zhao-xiang,MA Xiao-yan,ZHANG Wei.Development of a loop-mediated isothermal amplification forthe detection of Yersinia enterocolitica[J].Food Science and Technology,2011(9):335-339. |
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| Authors: | LIANG Lei LI Ying-jun MENG Zhao-xiang MA Xiao-yan ZHANG Wei |
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| Affiliation: | LIANG Lei,LI Ying-jun,MENG Zhao-xiang,MA Xiao-yan,ZHANG Wei*(College of Food Science and Technology,Agricultural University of Hebei,Baoding 071000) |
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| Abstract: | A loop-mediated isothermal amplification(LAMP) technology with two loop primers was developed for rapidly detecting Yersinia enterocolitica in chicken. Sequences of Ail gene of Y.e (AY004311.1) were used as target sequences, to design outer primers, inner primers. We judged the results of detection, through visible to the naked eye of the white precipitate. The result indicates: we extracted DNA using the method of Chemical reagent. The detection limit of pure bacterial culture was 6.3 cfu/mL and the detect... |
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| Keywords: | loop-mediated isothermal amplification detection Yersinia enterocolitica |
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