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
{"title":"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。","authors":"Mayasar I Al-Zaban,&nbsp;Ahlam H Alrokban,&nbsp;Mohamed A Mahmoud","doi":"10.1080/21501203.2023.2213704","DOIUrl":null,"url":null,"abstract":"<p><p>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 <i>Aspergillus</i> (4), <i>Fusarium</i> (3), <i>Penicillium</i> (3), and <i>Alternaria</i> (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 <i>Aspergillus, Fusarium, Penicillium</i>, and <i>Alternaria</i>. 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 (<i>aflQ, aflP, aflO</i>, and <i>aflD</i>), 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.</p>","PeriodicalId":18833,"journal":{"name":"Mycology","volume":"14 3","pages":"227-238"},"PeriodicalIF":4.6000,"publicationDate":"2023-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://ftp.ncbi.nlm.nih.gov/pub/pmc/oa_pdf/db/93/TMYC_14_2213704.PMC10424615.pdf","citationCount":"0","resultStr":"{\"title\":\"Development of a real-time PCR and multiplex PCR assay for the detection and identification of mycotoxigenic fungi in stored maize grains.\",\"authors\":\"Mayasar I Al-Zaban,&nbsp;Ahlam H Alrokban,&nbsp;Mohamed A Mahmoud\",\"doi\":\"10.1080/21501203.2023.2213704\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p><p>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 <i>Aspergillus</i> (4), <i>Fusarium</i> (3), <i>Penicillium</i> (3), and <i>Alternaria</i> (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 <i>Aspergillus, Fusarium, Penicillium</i>, and <i>Alternaria</i>. 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 (<i>aflQ, aflP, aflO</i>, and <i>aflD</i>), 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.</p>\",\"PeriodicalId\":18833,\"journal\":{\"name\":\"Mycology\",\"volume\":\"14 3\",\"pages\":\"227-238\"},\"PeriodicalIF\":4.6000,\"publicationDate\":\"2023-01-01\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"https://ftp.ncbi.nlm.nih.gov/pub/pmc/oa_pdf/db/93/TMYC_14_2213704.PMC10424615.pdf\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Mycology\",\"FirstCategoryId\":\"1085\",\"ListUrlMain\":\"https://doi.org/10.1080/21501203.2023.2213704\",\"RegionNum\":2,\"RegionCategory\":\"生物学\",\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"Q1\",\"JCRName\":\"MYCOLOGY\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Mycology","FirstCategoryId":"1085","ListUrlMain":"https://doi.org/10.1080/21501203.2023.2213704","RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q1","JCRName":"MYCOLOGY","Score":null,"Total":0}
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。

摘要图片

摘要图片

摘要图片

查看原文
分享 分享
微信好友 朋友圈 QQ好友 复制链接
本刊更多论文
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.

求助全文
通过发布文献求助,成功后即可免费获取论文全文。 去求助
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
Isolation of triazole-resistant Aspergillus fumigatus harbouring cyp51A mutations from five patients with invasive pulmonary aspergillosis in Yunnan, China Analytical and clinical validation of multiplex droplet digital PCR assay for detecting pathogenic fungal infection in lungs Optimisation of hypocrellin production in Shiraia -like fungi via genetic modification involving a transcription factor gene and a putative monooxygenase gene Call me by your name: Considerations of DNA sequences as types within wider discussions on fungal nomenclature A comprehensive review of secondary metabolites from the genus Agrocybe : Biological activities and pharmacological implications
×
引用
GB/T 7714-2015
复制
MLA
复制
APA
复制
导出至
BibTeX EndNote RefMan NoteFirst NoteExpress
×
×
提示
您的信息不完整,为了账户安全,请先补充。
现在去补充
×
提示
您因"违规操作"
具体请查看互助需知
我知道了
×
提示
现在去查看 取消
×
提示
确定
0
微信
客服QQ
Book学术公众号 扫码关注我们
反馈
×
意见反馈
请填写您的意见或建议
请填写您的手机或邮箱
已复制链接
已复制链接
快去分享给好友吧!
我知道了
×
扫码分享
扫码分享
Book学术官方微信
Book学术文献互助
Book学术文献互助群
群 号:481959085
Book学术
文献互助 智能选刊 最新文献 互助须知 联系我们:info@booksci.cn
Book学术提供免费学术资源搜索服务,方便国内外学者检索中英文文献。致力于提供最便捷和优质的服务体验。
Copyright © 2023 Book学术 All rights reserved.
ghs 京公网安备 11010802042870号 京ICP备2023020795号-1