李斯特氏菌因子血清的制备及食品检测的免疫磁性分离-PCR(MIPA)方法的建立  

Production of Factor Sera of Listeria and Establishment of the MIPA for Detection of Food

作　　者：商海涛[1] 柳增善[1] 陈贵连[1] 刘明远[1] 张让堂 徐克诚 王跟领[1] 

机构地区：[1]解放军农牧大学动物医学系,长春130062

出　　处：《中国兽医学报》1999年第4期339-343,共5页Chinese Journal of Veterinary Science

基　　金：军队青年基金

摘　　要：首先进行了李斯特氏菌因子血清的研制，制备出了可对所有李斯特氏菌分型的 １５ 个 Ｏ 抗原因子和４ 个 Ｈ 抗原因子的因子血清。利用复合因子血清的多克隆抗体包被磁性球，对食品中的单核细胞增多性李斯特氏菌进行免疫磁性分离，并与 Ｐ Ｃ Ｒ 方法相结合，建立了检测食品中单核细胞增多性李斯特氏菌的 Ｍ Ｉ Ｐ Ａ方法（免疫磁性分离—聚合酶链反应方法，ｍ ａｇｎ ｅｔｉｃ ｉｍ ｍ ｕｎｏｐｏｌｙｍ ｅｒａｓｅ ｃｈａｉｎ ｒｅａｃｔｉｏｎ ａｓｓａｙ）。对菌液、模拟样品的检测表明，本方法能够有效地克服食品基质、培养基成分和杂菌对 Ｐ Ｃ Ｒ 检验的干扰作用。食品样品在 Ｅ Ｂ增菌液中增菌 １２ ｈ 后，检测的敏感度达 ５ Ｃ Ｆ Ｕ／ｍ Ｌ，可以在 ２０ ｈ 内完成检测。本方法对实际食品样品的检测结果，与国家标准检验方法检测的结果一致。The factor sera of Listeria which can be used to types of all Listeria strains,including 15 O factors and 4 H factors were produced.Polycolonal antibodies in the multi factor sera were used to coat the magnetic beads to produce immunomagnetic beads isolating Listeria strains from foods,which was combined with polymerase chain reaction assay to establish the magnetic immuno polymerase chain reaction assay(MIPA).The detection of Listeria monocytogenes in cultures and food samples polluted artificially indicated that the method could overcome obstruction of food components, media components and other bacterium to PCR.The sensitivity of the MIPA or detection in food samples polluted artificially was 5 CFU/mL.The program of the MIPA can be finished in 20 hours.Listeria monocytogenes in actual foods were detected by the method,and the results accorded with that of the GB method.

关 键 词：单核细胞增多性 李斯特氏菌 食品 免疫磁性分离 

分 类 号：R512.701[医药卫生—内科学]