Mac Kiem, Wichmann-Schauer Heidi, Peters Jan, Ellerbroek Lüppo
Federal Institute of Consumer Health Protection and Veterinary Medicine, Diedersdorfer Weg 1, D-12277 Berlin, Germany.
Int J Food Microbiol. 2003 Dec 1;88(2-3):305-9. doi: 10.1016/s0168-1605(03)00192-2.
A multiplex PCR assay for the differentiation of species and detection of vancomycin (van) resistance genes in enterococci was established. Three hundred sixty-seven enterococcal strains isolated from food were selected from a sample collection and were examined with regards to the existence of four vancomycin resistance genes (vanA, vanB, vanC1 and vanC2) and to species differentiation (E. faecalis, E. faecium, E. casseliflavus and E. gallinarum) by multiplex PCR. Apart from unambiguous results for the majority of strains, the E. faecium specific primers used here, showed weak points in the specificity and sensitivity, respectively. Despite these rare instances, we succeeded in optimising the multiplex PCR assay by variation of annealing temperature and primer combination. The assay presented here, with its optimised parameters, is therefore suitable for routine use and because of time and material saving, it is an alternative and rapid method in comparison to conventional tests.
建立了一种用于肠球菌物种鉴别及检测万古霉素（van）耐药基因的多重聚合酶链反应（PCR）检测方法。从样本收集中选取367株从食品中分离出的肠球菌菌株，通过多重PCR检测四个万古霉素耐药基因（vanA、vanB、vanC1和vanC2）的存在情况以及进行物种鉴别（粪肠球菌、屎肠球菌、格氏肠球菌和鹑鸡肠球菌）。除了大多数菌株获得明确结果外，此处使用的屎肠球菌特异性引物分别在特异性和敏感性方面存在弱点。尽管有这些罕见情况，但我们通过改变退火温度和引物组合成功优化了多重PCR检测方法。因此，此处介绍的具有优化参数的检测方法适用于常规使用，并且由于节省时间和材料，与传统检测方法相比，它是一种替代的快速方法。
