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一种基于环介导等温扩增(LAMP)的快速检测产伏马菌素曲霉属物种的方法。

A loop-mediated isothermal amplification (LAMP) assay for rapid detection of fumonisin producing Aspergillus species.

机构信息

Institute of Sciences of Food Production (ISPA), National Research Council (CNR), Bari, Italy.

Institute of Sciences of Food Production (ISPA), National Research Council (CNR), Bari, Italy.

出版信息

Food Microbiol. 2020 Sep;90:103469. doi: 10.1016/j.fm.2020.103469. Epub 2020 Feb 28.

DOI:10.1016/j.fm.2020.103469
PMID:32336366
Abstract

Fumonisins contamination of food commodities is a worldwide problem, especially for maize. The ability to produce fumonisinsis a trait of several species of Fusarium, mainly F. verticillioides and F. proliferatum on maize, and some Aspergillus species. A. niger and its sister species A. welwitschiae, can contribute to fumonisin B (FB) accumulation in maize kernels, although to a lesser extent than fumonisin-producing Fusarium species. Fumonisins risk monitoring represents an effective strategy in the integrated approach for mycotoxin risk management and reduction. The availability of a user-friendlymolecular assay for the detection oftoxigenic fungal species represents a valuable tool in understanding and managing upcoming mycotoxin contamination. In this study, we developed a LAMP assay, based on the detection of fum10, for a rapid and specific molecular detection of FB-producing A. niger and A. welwistchiae, potentially useful to perform monitoring directly "on site" in maize chain. Results showed that very low amounts of conidia are suitable to detect the presence of the target gene, thus providing information about the presence of FB-producing Aspergillus species and the possible upcoming fumonisins contamination in maize. The assay was combined with a suitable protocol for "in field" crude DNA extraction and a colorimetric method for easy naked-eye evaluationof results, offering a reliable and user-friendly tool to support effective reduction strategies of mycotoxin contamination in crop management programs.

摘要

真菌毒素污染是一个全球性问题,特别是在玉米上。产生伏马菌素的能力是几种镰刀菌属物种的特征,主要是玉米上的尖孢镰刀菌和层出镰刀菌,以及一些曲霉菌属物种。黑曲霉及其姐妹种黄曲霉 welwitschiae 也可以导致玉米籽粒中伏马菌素 B(FB)的积累,尽管其积累程度低于产伏马菌素的镰刀菌属物种。真菌毒素风险监测是真菌毒素风险管理和降低综合方法中的有效策略。易于使用的分子检测方法可用于检测产毒真菌物种,这是了解和管理即将出现的真菌毒素污染的宝贵工具。在这项研究中,我们开发了一种基于 fum10 检测的 LAMP 检测方法,用于快速、特异性地检测产 FB 的黑曲霉和黄曲霉,这可能有助于在玉米链上直接“现场”进行监测。结果表明,非常少量的分生孢子即可检测到靶基因的存在,从而提供有关产 FB 的曲霉菌属物种存在以及玉米中可能出现伏马菌素污染的信息。该检测方法与适合现场粗提 DNA 提取的方案以及用于肉眼评估结果的比色法相结合,提供了一种可靠且易于使用的工具,可支持在作物管理计划中实施有效的真菌毒素污染减少策略。

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