利用定量实时 PCR 检测法可靠、特异地检测盘巾中的棘阿米巴属虫

IF 4.5 1区 农林科学 Q1 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Food microbiology Pub Date : 2024-05-18 DOI:10.1016/j.fm.2024.104562
M. Reyes-Batlle , E. Córdoba-Lanús , A. Domínguez-de-Barros , I. Sifaoui , R.L. Rodríguez-Expósito , S. Mantesa-Rodríguez , J.E. Piñero , J. Lorenzo-Morales
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引用次数: 0

摘要

阿卡阿米巴属(Acanthamoeba spp.)是一种无处不在的原生动物,属于自由生活阿米巴(FLA)类,被认为是造成副威胁性角膜炎或致命性脑炎等人类感染的病原体。此外,据报道,这种寄生虫还是对人类健康十分重要的其他微生物的宿主,如弯曲杆菌属或弧菌属等。棘阿米巴既是病原体,又是环境吞噬细胞,因此有越来越多的报告证实,棘阿米巴存在于与人类相关的环境中,是一种水质指标。考虑到水和厨房环境之间的潮汐关系,以及水源中棘阿米巴的高流行率,本研究旨在建立一个基于 DNA 提取和实时 qPCR 分析的快速、准确的方案,以检测洗碗布中的棘阿米巴属。通过处理 17 块使用过的洗碗布,验证了该程序的有效性。我们的研究结果表明,所使用的 qPCR 检测方法灵敏度很高,能够从体外污染的洗碗布中检测出最多一条棘阿米巴。该方法能准确检测出 64.7% 的阳性样本中的棘阿米巴属(4 个样本的 DNA 浓度相当于 1-102 个阿米巴)。我们的研究结果表明了通过高效灵敏的方法对 FLA 进行监测的重要性,因为这种阿米巴能在厨房水槽等与人类相关的食物环境中定植,并可能成为潜在的感染源。
本文章由计算机程序翻译,如有差异,请以英文原文为准。

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Reliable and specific detection of Acanthamoeba spp. In dishcloths using quantitative real-time PCR assay

Acanthamoeba spp., are ubiquitous protist which belongs to Free-Living Amoeba (FLA) group, is considered as causal agent of side-threatening keratitis or fatal encephalitis among other human infections. Besides, this parasite has been reported as host for other microorganisms important to human health such as Campylobacter spp. or Vibrio spp. among others. This role of Acanthamoeba as pathogen and environmental phagocyte has increased the reports confirming its presence in human related environments, acting as a water quality indicator. Considering the tide relationship between water and kitchen environments, and the high prevalence of Acanthamoeba in water sources, the present study aims to establish a quick and accurate protocol based on DNA extraction and a real time qPCR assay to detect Acanthamoeba spp. in dishcloths. The procedure has been validated by processing 17 used dishcloths. Our findings demonstrated the high sensitivity of the qPCR assay used which was capable of detecting up to one Acanthamoeba from an in vitro contaminated dishcloth. The protocol accurately detected 64.7% of positive samples for Acanthamoeba spp, (in 4 samples DNA concentrations corresponded to 1-102 amoebae). Our findings demonstrate the importance of FLA surveillance by efficient and sensitive methods since one amoeba is capable of colonizing human related food environments such as kitchens sinks and could be a potential source of infection.

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来源期刊
Food microbiology
Food microbiology 工程技术-生物工程与应用微生物
CiteScore
11.30
自引率
3.80%
发文量
179
审稿时长
44 days
期刊介绍: Food Microbiology publishes original research articles, short communications, review papers, letters, news items and book reviews dealing with all aspects of the microbiology of foods. The editors aim to publish manuscripts of the highest quality which are both relevant and applicable to the broad field covered by the journal. Studies must be novel, have a clear connection to food microbiology, and be of general interest to the international community of food microbiologists. The editors make every effort to ensure rapid and fair reviews, resulting in timely publication of accepted manuscripts.
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