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高通量免疫珠阵列技术用于鸡产品中三种食源性致病菌的多重检测的验证。

Validation of a high-throughput immunobead array technique for multiplex detection of three foodborne pathogens in chicken products.

机构信息

National Center for Genetic Engineering and Biotechnology (BIOTEC), National Science and Technology Development Agency (NSTDA), Pathum Thani, Thailand.

Institute for Global Food Security, School of Biological Sciences, Queen's University Belfast, Belfast, United Kingdom.

出版信息

Int J Food Microbiol. 2016 May 2;224:47-54. doi: 10.1016/j.ijfoodmicro.2016.02.017. Epub 2016 Feb 27.

Abstract

This study rigorously evaluated a previously developed immunobead array method to simultaneously detect three important foodborne pathogens, Campylobacter jejuni, Listeria monocytogenes, and Salmonella spp., for its actual application in routine food testing. Due to the limitation of the detection limit of the developed method, an enrichment step was included in this study by using Campylobacter Enrichment Broth for C. jejuni and Universal Pre-enrichment Broth for L. monocytogenes and Salmonella spp.. The findings showed that the immunobead array method was capable of detecting as low as 1CFU of the pathogens spiked in the culture media after being cultured for 24h for all three pathogens. The immunobead array method was further evaluated for its pathogen detection capabilities in ready-to-eat (RTE) and ready-to-cook (RTC) chicken samples and proven to be able to detect as low as 1CFU of the pathogens spiked in the food samples after being cultured for 24h in the case of Salmonella spp., and L. monocytogenes and 48 h in the case of C. jejuni. The method was subsequently validated with three types of chicken products (RTE, n=30; RTC, n=20; raw chicken, n=20) and was found to give the same results as the conventional plating method. Our findings demonstrated that the previously developed immunobead array method could be used for actual food testing with minimal enrichment period of only 52 h, whereas the conventional ISO protocols for the same pathogens take 90-144 h. The immunobead array was therefore an inexpensive, rapid and simple method for the food testing.

摘要

本研究严格评估了先前开发的免疫珠阵列方法,以同时检测三种重要的食源性病原体,即空肠弯曲菌、单核细胞增生李斯特菌和沙门氏菌,以将其实际应用于常规食品检测中。由于开发方法的检测限有限,本研究包括了一个富集步骤,分别使用空肠弯曲菌富集肉汤和通用预富集肉汤对空肠弯曲菌、单核细胞增生李斯特菌和沙门氏菌进行富集。研究结果表明,免疫珠阵列方法能够在所有三种病原体培养 24 小时后,检测到低至 1CFU 的病原体。该免疫珠阵列方法进一步用于评估其在即食(RTE)和即煮(RTC)鸡肉样品中的病原体检测能力,并证明在沙门氏菌培养 24 小时、单核细胞增生李斯特菌和空肠弯曲菌培养 48 小时后,能够检测到低至 1CFU 的病原体。该方法随后用三种类型的鸡肉产品(RTE,n=30;RTC,n=20;生鸡肉,n=20)进行了验证,结果与传统平板法相同。我们的研究结果表明,先前开发的免疫珠阵列方法可用于实际食品检测,仅需最短 52 小时的富集时间,而相同病原体的传统 ISO 方案则需要 90-144 小时。因此,免疫珠阵列是一种廉价、快速和简单的食品检测方法。

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