Herrera Fanny C, Santos Jesús A, Otero Andrés, García-López María-Luisa
Department of Food Hygiene and Food Technology, University of León, 24071-León, Spain.
Int J Food Microbiol. 2006 Apr 25;108(2):233-8. doi: 10.1016/j.ijfoodmicro.2005.12.008. Epub 2006 Feb 14.
PCR primers were designed and used to amplify a 435-bp fragment from the Plesiomonas shigelloides hugA gene. The PCR assay combined with a non-selective enrichment step proved to be a reliable procedure for P. shigelloides detection in fish meat. The incidence of this bacterium was investigated in 52 lots of pre-packed saltwater fish portions (conger, swordfish, sole, grouper, whiting and halibut) displayed at two hypermarkets by a conventional two-step procedure and the PCR assay. Using the former, P. shigelloides was isolated from three lots of grouper fillets and one lot of halibut fillets. When PCR was performed with non-selective enriched cultures of fish portions, amplification products were obtained from samples that were positive by the culturing method and from eight additional lots of grouper fillets that gave negative results with the conventional procedure. After a secondary enrichment in tetrathionate broth without iodine, all PCR-positive non-selective enrichments yielded P. shigelloides colonies. Overall, P. shigelloides was found in 23% of the examined lots of marine fish (11 of grouper and one of halibut).
设计了聚合酶链反应(PCR)引物,并用于扩增嗜水气单胞菌hugA基因的一个435碱基对的片段。PCR检测结合非选择性富集步骤被证明是检测鱼肉中嗜水气单胞菌的可靠方法。采用传统的两步法和PCR检测,对两家大型超市展示的52批预包装海水鱼块(海鳗、箭鱼、鳎鱼、石斑鱼、牙鳕和大比目鱼)中该细菌的发生率进行了调查。使用传统方法,从三批石斑鱼片和一批大比目鱼片分离出嗜水气单胞菌。当对鱼块的非选择性富集培养物进行PCR时,从培养方法呈阳性的样品以及另外八批传统方法检测结果为阴性的石斑鱼片中获得了扩增产物。在不含碘的连四硫酸盐肉汤中进行二次富集后,所有PCR阳性的非选择性富集培养物均产生了嗜水气单胞菌菌落。总体而言,在所检测的23%的海鱼批次(11批石斑鱼和1批大比目鱼)中发现了嗜水气单胞菌。