Impacts of phenolic compounds on RT-qPCR detection of hepatitis A virus in berries

IF 5.4 1区农林科学 Q1 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Food microbiology Pub Date : 2026-06-01 Epub Date: 2025-12-09 DOI:10.1016/j.fm.2025.105005

Marie-Ève Collard , Éric Jubinville , Valérie Goulet-Beaulieu , Julie Jean

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引用次数: 0

Abstract

Berries are frequently implicated in outbreaks of foodborne illness due to viruses, particularly norovirus and hepatitis A virus. Compounds naturally present in berries can compromise the reliability of RT-qPCR methods, such as ISO 15216–1:2017, for detecting and quantifying viruses in foods. The aim of this study was to evaluate the inhibitory impact of seven phenolic compounds (ellagic acid, hydroxybenzoic acid, caffeic acid, coumaric acid, ferulic acid, quercetin, and cyanidine-3-glucoside) found naturally in raspberries as well as batch effects due to different concentrations of inhibitors (e.g., associated with ripeness) when using RT-qPCR to detect HAV in raspberries, blackberries, strawberries, blueberries, cranberries, and mixed berries. To assess the impact of dilution on RT-qPCR inhibition, samples were diluted at four levels (1/2, 1/5, 1/10, 1/100). Spiking the RT-qPCR reaction mixture with each phenolic compound at its natural concentration in raspberries showed that ellagic acid, hydroxybenzoic acid, caffeic acid and cyanidin-3-glucoside inhibited amplification, but only ellagic acid remained inhibitory in the ISO method. HAV recovery from frozen strawberries was undetectable (0 %) without additional treatment but reached 39 % with the OneStep PCR Inhibitor Removal Kit. For frozen blueberries, MobiSpin S-400 performed better, yielding about 52 % recovery compared to 23 % with OneStep. Sample dilution further enhanced HAV detection across most berry types.

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酚类化合物对浆果中甲型肝炎病毒RT-qPCR检测的影响

浆果经常与病毒引起的食源性疾病暴发有关，特别是诺如病毒和甲型肝炎病毒。浆果中天然存在的化合物可能会影响RT-qPCR方法的可靠性，例如ISO 15216-1:2017，用于检测和定量食品中的病毒。本研究的目的是评估7种酚类化合物（鞣花酸、羟基苯甲酸、咖啡酸、香豆酸、阿魏酸、槲皮素和花青素-3-葡萄糖苷）在覆盆子中天然存在的抑制作用，以及使用RT-qPCR检测覆盆子、黑莓、草莓、蓝莓、蔓越莓和混合浆果中不同浓度抑制剂（如与成熟度相关的抑制剂）的批次效应。为了评估稀释对RT-qPCR抑制作用的影响，将样品稀释至4个水平（1/2、1/5、1/10、1/100）。用覆盆子中各酚类化合物的天然浓度进行RT-qPCR反应，发现鞣花酸、羟基苯甲酸、咖啡酸和花青素-3-葡萄糖苷抑制扩增，但只有鞣花酸在ISO法中保持抑制作用。在没有额外处理的情况下，冷冻草莓的HAV恢复率为0%，但使用OneStep PCR抑制剂去除试剂盒，HAV恢复率达到39%。对于冷冻蓝莓，MobiSpin S-400表现更好，回收率约为52%，而OneStep的回收率为23%。样品稀释进一步增强了大多数浆果类型的HAV检测。

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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来源期刊

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.