Gouws P A, Visser M, Brözel V S
Department of Microbiology, University of the Western Cape, Bellville, South Africa.
J Food Prot. 1998 Aug;61(8):1039-42. doi: 10.4315/0362-028x-61.8.1039.
Salmonella spp. are one of the most important groups of food-borne pathogens worldwide. Conventional methods for the detection of Salmonella spp. in foodstuffs are generally cumbersome and time consuming. Whereas various more rapid detection methods have been developed over the past few years, there is currently no reliable true 24-hour detection method available. We report here a reliable Salmonella PCR detection method yielding results within 24 h. Chicken samples were preenriched in buffered peptone water (BPW) for 6 h. The DNA was extracted using phosphate-buffered saline (PBS) and then heated at 95 degrees C for 10 min. The Salmonella-specific primers ST11 and ST15 were used to amplify a 429-bp region specific to all Salmonella spp. This approach proved to be sufficient for the reliable detection of Salmonella spp. from both artificially and naturally contaminated poultry samples. The characteristic 429-bp PCR product was obtained in artificially contaminated samples with a detection limit of 50 CFU. A variety of chicken samples confirmed to harbor Salmonella spp. by conventional culture methods tested positive by our 24-h procedure, whereas no detectable amplification product was detected in those samples testing negative by culture methods. This method proved to be an excellent tool for the rapid and sensitive detection of Salmonella spp. from poultry samples using a specific primer set (ST11 and ST15) after only 6 h of preenrichment.
沙门氏菌属是全球最重要的食源性病原体群体之一。食品中沙门氏菌属的传统检测方法通常繁琐且耗时。尽管在过去几年中已开发出各种更快速的检测方法，但目前尚无可靠的真正24小时检测方法。我们在此报告一种可靠的沙门氏菌PCR检测方法，可在24小时内得出结果。鸡肉样品在缓冲蛋白胨水（BPW）中预富集6小时。使用磷酸盐缓冲盐水（PBS）提取DNA，然后在95℃加热10分钟。沙门氏菌特异性引物ST11和ST15用于扩增所有沙门氏菌属特有的429bp区域。该方法被证明足以从人工和自然污染的家禽样品中可靠地检测出沙门氏菌属。在人工污染的样品中获得了特征性的429bp PCR产物，检测限为50 CFU。通过传统培养方法确认含有沙门氏菌属的各种鸡肉样品通过我们的24小时程序检测为阳性，而在那些通过培养方法检测为阴性的样品中未检测到可检测的扩增产物。该方法被证明是一种出色的工具，用于在仅预富集6小时后使用特定引物组（ST11和ST15）从家禽样品中快速灵敏地检测沙门氏菌属。
