九种单核细胞增生性李斯特菌检测技术效果比较及评价  

Comparison and Evaluation of Nine Methods in Detecting Listeria monocytogenes

作　　者：刘雅莉[1,2] 刘芳 韩舜愈[1] 刘箐[2] 孙志博[4] 夏俊芳[1] 朱小清 

机构地区：[1]甘肃农业大学食品科学与工程学院,兰州730070 [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,兰州730020 [4]兰州大学基础医学院,兰州730000 [5]甘肃出入境检验检疫局,兰州730020

出　　处：《中国生物工程杂志》2012年第6期84-92,共9页China Biotechnology

摘　　要：采用传统生物学方法、显色培养基方法、自制三抗体夹心ELISA试剂盒(TAS-ELISA)、基因组直接PCR(DNA Direct-PCR)、菌裂解直接PCR(Bacterium Direct-PCR)、免疫捕捉PCR(IC-PCR)、生物PCR(Bio-PCR)、SYBR Green染料实时荧光PCR(SYBR Green Real time-PCR)、探针实时荧光PCR(TaqMan Real time-PCR)检测纯菌液及模拟带菌食品中的单核细胞增生性李斯特菌(Listeria monocytogenes,LM)。结果表明:传统生物学培养方法出现假阳性;TAS-ELISA、DNADirect-PCR、Bacterium Direct-PCR、IC-PCR最低检测限分别为106、102、105、104CFU/ml;显色培养基、SYBR Green Real time-PCR灵敏度达102CFU/ml;Bio-PCR、TaqMan Real time-PCR检测灵敏度最高,均为101CFU/ml。显色培养基、TAS-ELISA操作简单,适合大量样品的初检;IC-PCR具有灵敏、特异、快速、经济的优点,特别适合大体积样品中微量病原的检测;Bio-PCR、TaqMan Real time-PCR灵敏度高、特异性好,适合阳性样品的复检以及科研使用。Nine methods were used to detect Listeria monocytogenes in bacterial suspensions and artificial contaminated food extracts, including traditional biological methods, chromogenic medium method, TAS-ELISA, DNA Direct-PCR, Bacterium Direct-PCR, IC-PCR, Bio-PCR, SYBR Green Real time-PCR and TaqMan Real time-PCR, and the results were analyzed and compared. The result showed that Traditional biological methods had false positive. The lowest detectable limits of TAS-ELISA, DNA Direct-PCR, Bacterium Direct-PCR and IC-PCR were 10^6、10^2、10^5、10^4CFU/ml. The detection sensitivity of chromogenic medium method, SYBR Green Real time-PCR was 102CFU/ml. Bio-PCR, TaqMan Real time-PCR had the highest sensitivity which could reach 101CFU/ml. Chromogenic medium and TAS-ELISA had the advantages of simple operation, and they were suitable for preliminary screening of large samples. IC-PCR was a sensitive, specific, rapid, economic technology, suitable for detecting micro-pathogens in large volume samples particularly. Bio-PCR and TaqMan Real time-PCR had high sensitivity and specificity, which adapted to recheck and research.

关 键 词：单核细胞增生性李斯特菌 三抗体夹心ELISA 免疫捕捉PCR 生物PCR 实时荧光 定量PCR 

分 类 号：Q819[生物学—生物工程]