九种单核细胞增生性李斯特菌检测技术效果比较及评价

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。

Comparison and Evaluation of Nine Methods in Detecting Listeria monocytogenes

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 106,102,105,104CFU/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.