Phagomagnetic separation in combination with real-time PCR assay for detection of Salmonella Enteritidis and Typhimurium in dairy products

IF 1.9 4区 农林科学 Q4 BIOTECHNOLOGY & APPLIED MICROBIOLOGY
Yifeng Ding, Zhiwei Li, Chenxi Huang, Yiming Zhang, Jia Wang, Xiaohong Wang
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引用次数: 0

Abstract

Salmonella Enteritidis (SE) and Salmonella Typhimurium (ST) are the most commonly reported serotypes detected for the occurrence of Salmonellosis through foodborne transmission in recent years. In this study, a phagomagnetic separation in combination with qPCR (PhMS-qPCR) based on phage T102 as a recognition element was developed for rapid enrichment and detection of Salmonella in dairy products. Phage T102 was coated onto magnetic beads for capturing Salmonella from samples through magnetic separation, with a capture efficiency of ~90% demonstrated. During the subsequent qPCR process, captured Salmonella was identified by detecting the OmpC gene, and specific sdf and STM4495 genes to further distinguish the serotypes of Salmonella Enteritidis (SE) and Salmonella Typhimurium (ST), respectively. The whole procedure can be performed in 2.5 h with a low detection limit of 10 CFU/mL in PBS. Subsequently, this detection strategy was successfully applied for the detection of Salmonella and serotype identification in spiked milk samples. This cell separation and detection system offers a promising alternative for rapid and accurate detection of Salmonella or Salmonella spp.

结合实时 PCR 检测乳制品中的肠炎沙门氏菌和鼠伤寒沙门氏菌的噬磁分离法
肠炎沙门氏菌(Salmonella Enteritidis,SE)和鼠伤寒沙门氏菌(Salmonella Typhimurium,ST)是近年来最常报道的通过食源性传播引发沙门氏菌病的血清型。本研究开发了一种以噬菌体 T102 为识别元件的噬菌体磁分离与 qPCR(PhMS-qPCR)相结合的方法,用于快速富集和检测乳制品中的沙门氏菌。将噬菌体 T102 涂在磁珠上,通过磁分离从样品中捕获沙门氏菌,捕获效率约为 90%。在随后的 qPCR 过程中,通过检测 OmpC 基因、特异的 sdf 和 STM4495 基因来识别捕获的沙门氏菌,从而进一步区分肠炎沙门氏菌(SE)和鼠伤寒沙门氏菌(ST)的血清型。整个过程可在 2.5 小时内完成,在 PBS 中的检测限低至 10 CFU/mL。随后,这种检测策略被成功应用于检测牛奶样品中的沙门氏菌和血清型鉴定。这种细胞分离和检测系统为快速准确地检测沙门氏菌或沙门氏菌属提供了一种很有前途的选择。
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来源期刊
Journal of Food Safety
Journal of Food Safety 工程技术-生物工程与应用微生物
CiteScore
5.30
自引率
0.00%
发文量
69
审稿时长
1 months
期刊介绍: The Journal of Food Safety emphasizes mechanistic studies involving inhibition, injury, and metabolism of food poisoning microorganisms, as well as the regulation of growth and toxin production in both model systems and complex food substrates. It also focuses on pathogens which cause food-borne illness, helping readers understand the factors affecting the initial detection of parasites, their development, transmission, and methods of control and destruction.
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