A Novel Fluorescence cross-priming Amplification Based on Universal Molecular Beacon for Rapid and Specific Detection of Salmonella enterica in food Samples

IF 2.6 3区农林科学 Q2 FOOD SCIENCE & TECHNOLOGY

Food Analytical Methods Pub Date : 2024-06-20 DOI:10.1007/s12161-024-02638-1

Wenfu Hou, Juan Du, Tingting Liu, Wensi Wang, Youwei Ai, Min Zhou, Hongxun Wang, Zhuo Wang

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Abstract

A methodology with rapidity and specificity is of great significance for the effective control and management of outbreaks caused by Salmonella enterica as it has presented an obvious threat to food safety and public health worldwide. In this study, we aimed to develop a tube-closed and real time cross-priming amplification assay for the rapid and sensitive detection of S.enterica and decrease the possibility of detecting false-positive signals derived from aerosol contamination or non-specific amplification caused by traditional CPA assay. The reaction system was optimized and the results showed that molecular beacon CPA (MB-CPA) method was highly specific for detection of S. enterica. The sensitivity of established assay was found to be 10 CFU/mL, 100 CFU/25 g, 10 CFU/25 g in pure culture, chicken sample without and with 6 h enrichment, respectively. And the sensitivity of MB-CPA was 10 times higher than that of real-time PCR. An application of MB-CPA assay was conducted with 78 naturally contaminated food samples to test its practicality. After an enrichment step at 37℃ for 6 h, the results showed 100% sensitivity and 100% specificity compared with standard culture-based method. Considering its rapidity, user-friendliness, cost-effectiveness, this MB-CPA assay will aid in the broader application in food industry for the detection of S. enterica in small or resource-limited food testing laboratories.

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基于通用分子信标的新型荧光交叉引物扩增技术，用于快速、特异地检测食品样本中的肠炎沙门氏菌

肠炎沙门氏菌对全球食品安全和公共卫生构成了明显的威胁，因此一种快速、特异的方法对有效控制和管理肠炎沙门氏菌引起的疫情爆发具有重要意义。本研究旨在开发一种闭管实时交叉引物扩增检测方法，用于快速灵敏地检测肠炎沙门氏菌，降低传统 CPA 检测方法因气溶胶污染或非特异性扩增而产生假阳性信号的可能性。对反应体系进行了优化，结果表明分子信标 CPA（MB-CPA）方法对检测肠道病毒具有高度特异性。在纯培养物、未富集和富集 6 小时的鸡肉样品中，所建立的检测灵敏度分别为 10 CFU/mL、100 CFU/25 g、10 CFU/25 g。MB-CPA 的灵敏度是实时 PCR 的 10 倍。为了测试 MB-CPA 检测法的实用性，我们对 78 份自然污染的食品样本进行了检测。与基于标准培养的方法相比，经过 37℃ 6 小时的富集步骤后，结果显示灵敏度为 100%，特异性为 100%。考虑到其快速性、用户友好性和成本效益，该 MB-CPA 检测方法将有助于在小型或资源有限的食品检测实验室中更广泛地应用于食品行业的肠道病毒检测。

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来源期刊

Food Analytical Methods 农林科学-食品科技

CiteScore

6.00

自引率

3.40%

发文量

244

审稿时长

3.1 months

期刊介绍： Food Analytical Methods publishes original articles, review articles, and notes on novel and/or state-of-the-art analytical methods or issues to be solved, as well as significant improvements or interesting applications to existing methods. These include analytical technology and methodology for food microbial contaminants, food chemistry and toxicology, food quality, food authenticity and food traceability. The journal covers fundamental and specific aspects of the development, optimization, and practical implementation in routine laboratories, and validation of food analytical methods for the monitoring of food safety and quality.