免疫捕捉PCR和直接PCR技术检测单核细胞增生性李斯特菌比较研究  

Detection of Listeria monocytogenes by IC-PCR and Direct PCR

作　　者：刘雅莉[1] 刘箐[2] 刘芳 韩舜愈[4] 王婧[4] 夏俊芳 汪小波 樊学军[6] 田绿波[6] 

机构地区：[1]兰州大学第二医院,甘肃兰州730030 [2]上海理工大学医疗器械与食品学院,上海200093 [3]甘肃出入境检验检疫局国际旅行卫生保健中心,甘肃兰州730020 [4]甘肃农业大学食品科学与工程学院,甘肃兰州730070 [5]昌吉州疾病控制预防中心,新疆昌吉831100 [6]四川出入境检验检疫局国际旅行卫生保健中心,四川成都610041

出　　处：《食品科学》2012年第20期243-248,共6页Food Science

基　　金：国家质量监督检验检疫总局科研项目(GSCIQ_2010IK220)

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。Immuno-capture PCR（IC-PCR）,established by integrating immunological and molecular biological techniques was compared with traditional direct PCR.The sensitivity（104CFU/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,106CFU/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,ICPCR 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.

关 键 词：单核细胞增生性李斯特菌 免疫捕捉PCR 直接PCR 

分 类 号：TS207.4[轻工技术与工程—食品科学]