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Enterotoxigenicity and Genetic Relatedness of Clostridium perfringens Isolates from Retail Foods in the United States

机译:美国零售食品中产气荚膜梭状芽孢杆菌分离株的肠毒力和遗传相关性

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Clostridium perfringens is a leading cause of bacterial food-borne illness in countries where consumption of meat and poultry is high. For example, each year in the United States, this organism is the second or third most common cause of confirmed cases of food-borne illness. Surveys of the incidence of this organism in retail foods were done in the 1960s without regard to whether isolates were enterotoxigenic. It is now known that not all strains of this organism possess the enterotoxin gene responsible for illness. We examined the incidence of this organism in 131 food samples from retail food stores in an area of the northeastern United States. Forty isolates were obtained by using the iron milk method at 45°C, with confirmation by use of motility nitrate and lactose gelatin media. The presence of the C. perfringens enterotoxin (cpe) and alpha toxin (cpa) genes was determined by PCR using previously published primer sequences. All isolates possessed cpa. None of the isolates were identified as carrying the cpe gene by this method or by another method using a digoxigenin-labeled gene probe. Consistent with these results, none of the sporulating-cell extracts contained enterotoxin as determined by reverse passive latex hemagglutination. Pulsed-field gel electrophoresis was used to determine the genetic relatedness of the isolates. About 5% of the isolates were considered to be closely related (2- to 3-band difference). The others were considered to be unrelated to one another. The results demonstrate the rarity of cpe+ strains in retail foods and the genetic diversity among nonoutbreak strains.
机译:在肉类和家禽消费量很高的国家,产气荚膜梭菌是细菌性食源性疾病的主要原因。例如,在美国,每年这种生物是确诊食源性疾病的第二或第三大最常见原因。在1960年代对零售食品中这种生物的发生率进行了调查,而没有考虑到分离物是否具有肠毒性。现在已知并非该生物的所有菌株都具有引起疾病的肠毒素基因。我们在美国东北部地区的一家零售食品商店的131个食品样本中检查了这种生物的发生率。通过在45℃下使用铁乳法获得40种分离物,并通过使用硝酸盐和乳糖明胶培养基确认。产气荚膜梭菌肠毒素(cpe)和α毒素(cpa)基因的存在通过PCR使用先前公开的引物序列来确定。所有分离株均具有cpa。通过这种方法或使用洋地黄毒苷标记的基因探针的另一种方法,没有分离物被鉴定为携带cpe基因。与这些结果一致,通过反向被动乳胶血凝测定,孢子形成细胞提取物中均不含肠毒素。脉冲场凝胶电泳用于确定分离物的遗传相关性。大约5%的分离株被认为是密切相关的(2-3波段差异)。其他被认为是彼此无关的。结果表明,零售食品中cpe +菌株稀有,非暴发菌株之间的遗传多样性也不同。

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