| 摘要: |
| 【目的】确定适用于不同条件下快速检测单增李斯特氏菌的方法。【方法】以单增李斯特氏菌标准菌株CMCC54004和市售盐水鸭、凉拌菜为研究对象，按照国家食品安全标准（GB 4789.30-2010）对检测样品进行选择性增菌培养，选择阴性样品进行人工布菌，比较单增李斯特氏菌蛋白条检测法、CPA恒温扩增法以及实时荧光PCR方法的检测效果，并以传统平板分离法进行验证。【结果】蛋白条检测法操作简单但灵敏度低，最低检测限为106 cfu/mL；实时荧光PCR方法灵敏度高，最低检测限为101 cfu/mL,但操作繁琐，对操作技术及实验室条件要求较高；CPA恒温扩增方法灵敏度相对较高，最低检测限为105 cfu/mL,操作相对简单。【结论】实时荧光PCR方法适合灵敏度要求高、且具备良好实验条件时使用；CPA恒温扩增法适于对灵敏度要求不高、实验室条件简单的基层实验室使用。 |
| 关键词: 单增李斯特氏菌 检测方法比较 蛋白条检测法 CPA恒温扩增法 实时荧光PCR方法 |
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| 基金项目:国家质检总局课题(2012IK168)；现代农业产业技术体系专项 (CARS-42-G22)；动物源食品安全加工科技工程项目(2012BAD28B00)；青奥会食品安全保障关键技术应用与示范项目（2011BAK21B05） |
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| Comparison of three analytical methods for rapid detection of Listeria monocytogenes |
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ZHOU Yang,ZHU Chang-qing,GUO Gui-ping,et al
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| Abstract: |
| 【Objective】This study aimed to determine the best method for rapid detection of Listeria monocytogenes in different conditions.【Method】Taking the Listeria monocytogenes standard strain CMCC54004,commercial salted duck,and salad as research objects,selectively enrichment of detection samples were conducted and artificially contaminated negative samples according to national standards.Detection results of L.monocytogenes protein detection method,CPA isothermal amplification method,and real-time fluorescence PCR method were compared and verified with traditional flat separation process.【Result】Protein detection method was simple in operation with low sensitivity of 106 cfu/mL.Real-time fluorescence PCR method had high sensitivity of 101 cfu/mL,but it was cumbersome to operate and required high operation techniques and laboratory conditions.CPA isothermal amplification method had relatively high sensitivity of 105 cfu/mL and simple operation requirement.【Conclusion】The real-time fluorescent PCR-method was suitable for high sensitivity with good experimental conditions.CPA isothermal amplification method was suitable for low-sensitivity requirement and simple laboratory conditions. |
| Key words: Listeria monocytogenes comparison of test methods protein detection method CPA isothermal amplification method real-time fluorescent PCR method |
