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用于从食品基质中8小时检测的靶向基因的新型TaqMan实时荧光定量PCR。

Novel TaqMan real-time PCR targeting gene for 8-h detection of from food matrices.

作者信息

Isaac Ann Mary, Kolla Harish Babu, Pallavi K P, Bandyadka Sharat, Mhatre Chinmayi Nandkishor, Urs Radhika Madan, Kingston J Joseph

机构信息

Defence Institute of Biodefence Technologies (DIBT-DRDO), Mysore, India.

出版信息

Front Microbiol. 2025 Jul 16;16:1517680. doi: 10.3389/fmicb.2025.1517680. eCollection 2025.

Abstract

, a well-known food-borne zoonotic pathogen, is the causative agent of Salmonellosis affecting public health in both developed and developing countries. Traditional detection methods are time-consuming, involving multiple steps like pre-enrichment, selective plating, and biochemical confirmation. In this study, a faster and sensitive TaqMan real-time PCR assay was developed with a 6 h enrichment paradigm for the direct detection of from food matrices using a novel target gene, , which is part of the Type 3 Secretion System (T3SS). The assay was found to be highly specific and had a limit of detection of 10 CFU/mL in pure culture. With a 6-h enrichment step, the assay was able to detect even low bacterial inoculum like 10 CFU/mL of Typhimurium in both artificially contaminated raw foods (raw egg and frozen chicken) and cooked Indian food matrices (ready-to-eat chicken biriyani and chicken pulao). The assay demonstrated 100% relative sensitivity, specificity, and accuracy across 52 natural (raw and processed) food samples. In summary, the real-time detection method developed is faster, specific, sensitive and a potential tool for regular monitoring in diverse food matrices within 8 h.

摘要

作为一种著名的食源性人畜共患病原体,是沙门氏菌病的病原体,在发达国家和发展中国家均影响公众健康。传统检测方法耗时较长,涉及预富集、选择性平板接种和生化确认等多个步骤。在本研究中,开发了一种更快且灵敏的TaqMan实时PCR检测方法,采用6小时富集模式,利用一种新型靶基因(作为3型分泌系统(T3SS)的一部分)直接从食品基质中检测。该检测方法具有高度特异性,在纯培养物中的检测限为10 CFU/mL。经过6小时的富集步骤,该检测方法能够在人工污染的生食(生鸡蛋和冷冻鸡肉)和熟制印度食品基质(即食鸡肉比尔亚尼和鸡肉抓饭)中检测到低至10 CFU/mL的鼠伤寒沙门氏菌等低细菌接种量。该检测方法在52份天然(生的和加工过的)食品样本中显示出100%的相对灵敏度、特异性和准确性。总之,所开发的实时检测方法更快、特异、灵敏,是在8小时内对多种食品基质进行常规监测的潜在工具。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a4a7/12307383/2ba138974e89/fmicb-16-1517680-g001.jpg

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