Park S H, Ricke S C
Cell and Molecular Biology Program, Department of Food Science, University of Arkansas, Fayetteville, AR, USA; Center for Food Safety, Department of Food Science, University of Arkansas, Fayetteville, AR, USA.
J Appl Microbiol. 2015 Jan;118(1):152-60. doi: 10.1111/jam.12678. Epub 2014 Nov 21.
The aim of this research was to develop multiplex PCR assay that could simultaneously detect Salmonella genus, Salmonella subsp. I, Salm. Enteritidis, Heidelberg and Typhimurium because these Salmonella serovars are the most common isolates associated with poultry products.
Five primers were utilized to establish multiplex PCR and applied to Salmonella isolates from chickens and farm environments. These isolates were identified as Salmonella subsp. I and 16 of 66 isolates were classified as Salm. Enteritidis, while Heidelberg or Typhimurium was not detected. We also spiked three Salmonella strains on chicken breast meat to evaluate the specificity and sensitivity of multiplex PCR as well as qPCR to optimize quantification of Salmonella in these samples. The optimized multiplex PCR and qPCR could detect approx. 2·2 CFU of Salmonella per gram after 18 h enrichment.
The multiplex PCR and qPCR would provide rapid and consistent results. Also, these techniques would be useful for the detection and quantification of Salmonella in contaminated poultry, foods and environmental samples.
The strategy for the rapid detection of Salmonella serovars in poultry is needed to further reduce the incidence of salmonellosis in humans. The optimized multiplex PCR will be useful to detect prevalent Salmonella serovars in poultry products.
本研究旨在开发一种多重聚合酶链反应（PCR）检测方法，能够同时检测沙门氏菌属、沙门氏菌亚种I、肠炎沙门氏菌、海德堡沙门氏菌和鼠伤寒沙门氏菌，因为这些沙门氏菌血清型是与家禽产品相关的最常见分离株。
使用5种引物建立多重PCR，并应用于从鸡和养殖环境中分离出的沙门氏菌。这些分离株被鉴定为沙门氏菌亚种I，66株分离株中有16株被归类为肠炎沙门氏菌，未检测到海德堡沙门氏菌或鼠伤寒沙门氏菌。我们还在鸡胸肉上接种了三种沙门氏菌菌株，以评估多重PCR以及定量PCR（qPCR）的特异性和灵敏度，从而优化这些样品中沙门氏菌的定量分析。优化后的多重PCR和qPCR在富集18小时后，每克样品中可检测到约2.2个沙门氏菌菌落形成单位（CFU）。
多重PCR和qPCR能够提供快速且一致的结果。此外，这些技术对于检测和定量受污染家禽、食品及环境样品中的沙门氏菌将很有用。
需要采用快速检测家禽中沙门氏菌血清型的策略，以进一步降低人类沙门氏菌病的发病率。优化后的多重PCR将有助于检测家禽产品中常见的沙门氏菌血清型。
