Leidreiter Melanie, Kühne Michael
Landesamt für Verbraucherschutz und Lebensmittelsicherheit Niedersachsen, Lebensmittelinstitut Oldenburg.
Berl Munch Tierarztl Wochenschr. 2007 Jan-Feb;120(1-2):79-85.
A qualitative detection of Listeria monocytogenes was performed in spiked minced pork meat using ELISA, Multiplex-PCR, Microarray and cultural reference method. Minced pork meat in batches of 10 or 25 g was spiked with 25 cfu Listeria monocytogenes per gram and incubated in selective enrichment solutions. During enrichment samples were collected continiously and a Listeria monocytogenes-ELISA, a Multiplex-PCR with electrophoretical detection (Listeria duplex) and a PCR with detection by Microarray (NUTRI Chip) were performed. The enrichment time after which all sub-samples were positive was defined as minimum enrichment time. With the cultural reference method Listeria monocytogenes was detected in 100 % of the samples after a total analysis time of 5 days. With the ELISA kit used in this study positive results were achieved after enrichment for 24 h. Multiplex-PCR with electrophoretical detection was positive after only 16 h of enrichment. The most sensitive detection method was the microarray. Using this technique, an enrichment time of 10 h was sufficient to get positive results in all samples.
采用酶联免疫吸附测定（ELISA）、多重聚合酶链反应（Multiplex-PCR）、微阵列技术及传统培养法对添加了单核细胞增生李斯特菌的绞碎猪肉进行定性检测。将10克或25克批次的绞碎猪肉每克添加25个菌落形成单位（cfu）的单核细胞增生李斯特菌，并在选择性增菌液中培养。在增菌过程中持续采集样本，并进行单核细胞增生李斯特菌ELISA检测、带电泳检测的多重聚合酶链反应（李斯特菌双联检测）以及带微阵列检测的聚合酶链反应（营养芯片）。将所有子样本均呈阳性的增菌时间定义为最短增菌时间。采用传统培养法，在5天的总分析时间后，100%的样本中检测到了单核细胞增生李斯特菌。使用本研究中所用的ELISA试剂盒，增菌24小时后获得阳性结果。带电泳检测的多重聚合酶链反应在增菌仅16小时后呈阳性。最灵敏的检测方法是微阵列技术。使用该技术，10小时的增菌时间足以使所有样本获得阳性结果。
