Department of Food Quality and Safety, Institute for Postharvest and Food Sciences, Agricultural Research Organization, The Volcani Center, Rishon LeZion 7528809, Israel.
National Residue Control Laboratory, Kimron Veterinary Institute, Bet Dagan 50250, Israel.
Toxins (Basel). 2017 Sep 25;9(10):302. doi: 10.3390/toxins9100302.
This study aimed to assess the occurrence of toxigenic fungi and mycotoxin contamination in stored wheat grains by using advanced molecular and analytical techniques. A multiplex polymerase chain reaction (PCR) strategy was established for rapid identification of mycotoxigenic fungi, and an improved analytical method was developed for simultaneous multi-mycotoxin determination in wheat grains by liquid chromatography-tandem mass spectrometry (LC/MS/MS) without the need for any clean-up. The optimized multiplex PCR method was highly specific in detecting fungal species containing species-specific and mycotoxin metabolic pathway genes. The method was applied for evaluation of 34 wheat grain samples collected from storage warehouses for the presence of mycotoxin-producing fungi, and a few samples were found positive for and species. Further chemical analysis revealed that 17 samples contained mycotoxins above the level of detection, but only six samples were found to be contaminated over the EU regulatory limits with at least one mycotoxin. Aflatoxin B₁, fumonisins, and deoxynivalenol were the most common toxins found in these samples. The results showed a strong correlation between the presence of mycotoxin biosynthesis genes as analyzed by multiplex PCR and mycotoxin detection by LC/MS/MS. The present findings indicate that a combined approach might provide rapid, accurate, and sensitive detection of mycotoxigenic species and mycotoxins in wheat grains.
本研究旨在利用先进的分子和分析技术评估储存小麦中产毒真菌和霉菌毒素污染的情况。建立了一种多重聚合酶链反应（PCR）策略，用于快速鉴定产毒真菌，并开发了一种改进的分析方法，通过液相色谱-串联质谱（LC/MS/MS）无需任何净化即可同时测定小麦中的多种霉菌毒素。优化的多重 PCR 方法在检测含有种特异性和霉菌毒素代谢途径基因的真菌物种方面具有高度特异性。该方法用于评估从储存仓库采集的 34 个小麦样品中是否存在产毒真菌，发现少数样品呈阳性，存在 和 物种。进一步的化学分析表明，17 个样品中的霉菌毒素含量高于检测水平，但只有 6 个样品的至少一种霉菌毒素含量超过欧盟法规限量。在这些样品中，最常见的毒素是黄曲霉毒素 B₁、伏马菌素和脱氧雪腐镰刀菌烯醇。研究结果表明，通过多重 PCR 分析的霉菌生物合成基因的存在与 LC/MS/MS 检测的霉菌毒素之间存在很强的相关性。本研究结果表明，联合方法可能为小麦中产毒物种和霉菌毒素的快速、准确和敏感检测提供依据。
