RF-LAMP技术检测鸡肉中小肠结肠炎耶尔森氏菌  

Detection of Yersinia enterocolitica in Chicken by Real-Time Fluorescence Loop-Mediated Isothermal Amplification Method

作　　者：张蕴哲[1] 杨倩 马晓燕[1] 杨澜 张伟[1,2] 

机构地区：[1]河北农业大学食品科技学院,河北保定071000 [2]河北农业大学理工学院,河北沧州061100

出　　处：《食品研究与开发》2018年第4期133-138,共6页Food Research and Development

基　　金：河北农业大学引进博士专项(ZD201623);河北省高等学校科学技术研究重点项目(ZD2017237);河北省自然科学基金青年科学基金项目(C2017204027)

摘　　要：本研究建立实时荧光环介导等温扩增技术检测小肠结肠炎耶尔森氏菌的方法,扩增产物经电泳和酶切鉴定,表明扩增结果正确。本研究验证该方法特异性,对灵敏度和人工污染鸡肉的检出限进行测定。结果表明小肠结肠炎耶尔森氏菌株呈阳性结果,非小肠结肠炎耶尔森氏菌株均呈阴性结果。该方法检测纯菌的灵敏度为61 CFU/m L,人工污染鸡肉的检出限为46 CFU/g。该方法特异性强、灵敏度高、可实时检测小肠结肠炎耶尔森氏菌,能够实现对小肠结肠炎耶尔森氏菌的快速检测。This study aimed to establish a real-time fluorescence loop-mediated isothermal amplification（RF-LAMP）assay to detect Yersinia enterocolitica （Y. enterocolitica） . The products were identified by restrictiondigestion followed by electrophoresis and the results indicated that the amplification was correct. The performanceof the assay was evaluated with respect to specificity and sensitivity,and the detection limit of artificiallycontaminated chicken was tested. Among the strains tested,Y. enterocolitica strain was identified as positive,the other non-Y. enterocolitica strains were negative. The sensitivity for Y. enterocolitica was 61 CFU/mL inpure cultures. The detection limit for artificially contaminated chicken was 46 CFU/g. This method has highspecificity and sensitivity and allowed real-time monitoring,which was useful to realize rapid detection ofY . enterocolitica.

关 键 词：实时荧光 环介导等温扩增 小肠结肠炎耶尔森氏菌 检测 鸡肉 

分 类 号：O657.3[理学—分析化学] TS251.7[理学—化学]