Cocolin Luca, Rantsiou Kalliopi, Iacumin Lucilla, Cantoni Carlo, Comi Giuseppe
Dipartimento Scienze degli Alimenti, Università degli studi di Udine, Italy.
Appl Environ Microbiol. 2002 Dec;68(12):6273-82. doi: 10.1128/AEM.68.12.6273-6282.2002.
A new molecular approach for the detection and identification of Listeria spp. and Listeria monocytogenes in food is presented here. The method is based on the PCR amplification of a fragment of the iap gene from the five species belonging to the genus and on the analysis of the PCR products obtained by denaturing gradient gel electrophoresis (DGGE). The protocol was first optimized by using strains from international collections. Based on the differences present in the sequences amplified, it was possible to obtain species-specific DGGE migration that allowed fast and easy identification of L. monocytogenes, L. innocua, L. welshimeri, L. seeligeri, and L. ivanovii. Moreover, for L. monocytogenes serotypes, partial differentiation was possible. The optimized protocol was used for identification of Listeria strains traditionally isolated from food and for direct detection and identification of Listeria members in food after an overnight enrichment. Identification of 48 food isolates and direct detection of Listeria spp. in 73 food samples show the potential of the method that can be used as a fast screening test to investigate the presence of Listeria spp. and L. monocytogenes in food.
本文介绍了一种用于检测和鉴定食品中李斯特菌属及单核细胞增生李斯特菌的新分子方法。该方法基于对该属五个物种的iap基因片段进行PCR扩增,并对通过变性梯度凝胶电泳(DGGE)获得的PCR产物进行分析。该方案首先通过使用来自国际菌种保藏中心的菌株进行优化。基于扩增序列中存在的差异,能够获得物种特异性的DGGE迁移图谱,从而快速、简便地鉴定单核细胞增生李斯特菌、无害李斯特菌、威氏李斯特菌、斯氏李斯特菌和伊氏李斯特菌。此外,对于单核细胞增生李斯特菌的血清型,也能够进行部分区分。优化后的方案用于鉴定传统上从食品中分离出的李斯特菌菌株,以及在过夜增菌后直接检测和鉴定食品中的李斯特菌属成员。对48株食品分离株的鉴定以及对73份食品样品中李斯特菌属的直接检测,显示了该方法的潜力,可作为一种快速筛选试验来调查食品中李斯特菌属和单核细胞增生李斯特菌的存在情况。