建立实时荧光定量PCR和多重荧光定量PCR检测和鉴定玉米储藏中产霉菌毒素真菌的方法。

IF 4.6 2区 生物学 Q1 MYCOLOGY Mycology Pub Date : 2023-01-01 DOI:10.1080/21501203.2023.2213704
Mayasar I Al-Zaban, Ahlam H Alrokban, Mohamed A Mahmoud
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引用次数: 0

摘要

本研究旨在鉴定重要的产霉菌毒素真菌,并利用分子方法对贮藏玉米籽粒中的霉菌毒素进行准确检测。本研究还优化了实时PCR (RT-PCR)检测。建立熔解曲线,对分离真菌曲霉(Aspergillus, 4)、镰刀菌(Fusarium, 3)、青霉(Penicillium, 3)和Alternaria(1)进行鉴定。建立了一种多重聚合酶链反应(mPCR)技术,用于检测和鉴定产生霉菌毒素的真菌、霉菌毒素代谢途径基因,并利用高效液相色谱(HPLC)测定贮藏玉米籽粒中的11种霉菌毒素。mPCR结果显示,曲霉、镰刀菌、青霉和互交菌的潜在产霉毒素真菌种类检测呈阳性。采用多重逆转录聚合酶链反应(mRT-PCR)技术对游离和受黄曲霉毒素B1 (AFB1)污染的玉米进行了检测。4个黄曲霉毒素生物合成途径基因AFB1 (aflQ、aflP、aflO和aflD)的表达模式是玉米籽粒污染和贮藏的良好标记。HPLC分析结果显示,玉米籽粒样品中霉菌毒素超标。结果表明,多相法可为贮藏玉米籽粒中产霉菌毒素的真菌种类和真菌毒素的检测和鉴定提供一种灵敏、快速、准确的方法。
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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Development of a real-time PCR and multiplex PCR assay for the detection and identification of mycotoxigenic fungi in stored maize grains.

This study aimed to identify important mycotoxigenic fungi and accurate detection of mycotoxin in stored maize grains using molecular methods. The current study also optimised the real-time PCR (RT-PCR) assay. The melting curve was established to identify isolated fungal species of Aspergillus (4), Fusarium (3), Penicillium (3), and Alternaria (one). A multiplex polymerase chain reaction (mPCR) technique was developed for the detection and characterisation of mycotoxin producing fungi, mycotoxin metabolic pathway genes, and the determination of eleven mycotoxins in stored maize grains using high-performance liquid chromatography (HPLC). The mPCR results indicated positive signals for potentially mycotoxigenic fungal species tested of Aspergillus, Fusarium, Penicillium, and Alternaria. A protocol for multiplex reverse transcription-polymerase chain reaction (mRT-PCR) was tested to distinguish between free and contaminated, stored maize with aflatoxin B1 (AFB1). The expression pattern of four aflatoxin biosynthetic pathway genes, AFB1 (aflQ, aflP, aflO, and aflD), was a good marker for contaminated, stored maize grains. HPLC analysis showed that maize grain samples were contaminated with mycotoxins, and the concentration was above the detection level. The results indicate that the polyphasic approach might provide a sensitive, rapid, and accurate method for detecting and identifying mycotoxigenic fungal species and mycotoxins in stored maize grains.

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来源期刊
Mycology
Mycology Medicine-Infectious Diseases
CiteScore
9.10
自引率
0.00%
发文量
18
审稿时长
13 weeks
期刊最新文献
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