Jakociune Dziuginta, Pasquali Frédérique, da Silva Cristiana Soares, Löfström Charlotta, Hoorfar Jeffrey, Klein Günter, Manfreda Gerardo, Olsen John Elmerdahl
Department of Veterinary Disease Biology, Faculty of Health and Medical Sciences, University of Copenhagen, Frederiksberg, Denmark.
Appl Environ Microbiol. 2014 Mar;80(5):1616-22. doi: 10.1128/AEM.03360-13. Epub 2013 Dec 20.
Salmonellae are a major cause of food-borne outbreaks in Europe, with eggs and egg products being identified as major sources. Due to the low levels of salmonellae in eggs and egg products, direct quantification is difficult. In the present study, enrichment quantitative real-time PCR (qPCR) was employed for enumeration of salmonellae in different matrices: table eggs, pasteurized egg products, and egg-containing dishes. Salmonella enterica serovar Enteritidis and S. enterica serovar Tennessee were used to artificially contaminate these matrices. The results showed a linear regression between the numbers of salmonellae and the quantification cycle (Cq) values for all matrices used, with the exception of pasteurized egg white. Standard curves were constructed by using both stationary-phase cells and heat-stressed cells, with similar results. Finally, this method was used to evaluate the fate of salmonellae in two egg-containing dishes, long egg and tiramisu, at abused refrigeration temperatures, and results indicated the growth of bacteria over a 1-week period. In conclusion, enrichment qPCR was shown to be reliable for enumeration of salmonellae in different egg products.
沙门氏菌是欧洲食源性疾病爆发的主要原因,鸡蛋和蛋制品被确定为主要来源。由于鸡蛋和蛋制品中沙门氏菌含量较低,直接定量较为困难。在本研究中,采用富集定量实时荧光定量聚合酶链反应(qPCR)对不同基质中的沙门氏菌进行计数:鲜鸡蛋、巴氏杀菌蛋制品和含蛋菜肴。肠炎沙门氏菌肠炎血清型和肠炎沙门氏菌田纳西血清型被用于人工污染这些基质。结果表明,除巴氏杀菌蛋清外,所有使用的基质中沙门氏菌数量与定量循环(Cq)值之间均呈线性回归。使用稳定期细胞和热应激细胞构建标准曲线,结果相似。最后,该方法用于评估两种含蛋菜肴(卤蛋和提拉米苏)在冷藏温度被滥用的情况下沙门氏菌的变化情况,结果表明细菌在1周内生长。总之,富集qPCR被证明在计数不同蛋制品中的沙门氏菌时是可靠的。