食品科学 ›› 2012, Vol. 33 ›› Issue (20): 243-248.

• 分析检测 • 上一篇    下一篇

免疫捕捉PCR和直接PCR技术检测单核细胞增生性李斯特菌的比较研究

刘雅莉1,刘 箐2,*,刘 芳3,韩舜愈4,王 婧4,夏俊芳5,汪小波3,樊学军6,田绿波6   

  1. 1.兰州大学第二医院 2.上海理工大学医疗器械与食品学院 3.甘肃出入境检验检疫局国际旅行卫生保健中心 4.甘肃农业大学食品科学与工程学院 5.昌吉州疾病控制预防中心 6.四川出入境检验检疫局国际旅行卫生保健中心
  • 收稿日期:2011-09-21 修回日期:2012-09-26 出版日期:2012-10-25 发布日期:2012-11-09
  • 通讯作者: 刘箐 E-mail:liuq@usst.edu.cn
  • 基金资助:

    快速检测食源性致病菌蛋白芯片

Detection of Listeria monocytogenes by IC-PCR and Direct PCR

2   

  • Received:2011-09-21 Revised:2012-09-26 Online:2012-10-25 Published:2012-11-09

摘要: 将免疫学技术与分子生物学技术相结合,建立了免疫捕捉PCR(IC-PCR),并与传统的直接PCR(Direct-PCR)进行比较。结果显示:纯菌液中IC-PCR检测灵敏度(104CFU/mL)是Direct-PCR灵敏度(105CFU/mL)的10倍;模拟带菌实验结果表明:IC-PCR最低可检测到5×101个细菌/PCR反应体系(即104CFU/mL),检测限比Direct-PCR(5×103个细菌/PCR反应体系即106CFU/mL)高100倍;特异性实验、干扰实验结果表明IC-PCR可特异检测单核细胞增生性李斯特菌(Listeria monaytogenes,LM),而和18株其他常见食源性致病菌无交叉反应。IC-PCR具有快速、经济、灵敏、特异等优点,是一种适合食品安全监管部门、食品企业的LM快速检测方法。

关键词: 单核细胞增生性李斯特菌, 免疫捕捉PCR, 直接PCR

Abstract: Immuno-capture PCR (IC-PCR), established by integrating immunological and molecular biological techniques was compared with traditional direct PCR. The sensitivity (104 CFU/mL) of IC-PCR for pure Listeria monocytogenes culture suspensions indicated a 10-fold increase compared with direct PCR (104 CFU/mL). The limit of detection of IC-PCR was 5 × 101 bacteria/PCR reaction system (i.e. 104 CFU/mL), which was 100 times higher than that of direct PCR (5 × 103 bacteria/PCR reaction system, 106 CFU/mL). IC-PCR had high specificity and anti-interference capability for detecting Listeria monocytogenes. Moreover, no cross-reactivity with 18 common foodborne pathogens was observed. Therefore, IC-PCR is a rapid, economical, sensitive and specific so that it can be suitable for rapid diction of Listeria monocytogenes in food safety supervisory agencies and food enterprises.

Key words: Listeria monocytogenes, IC-PCR, direct PCR

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