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Development of real-time PCR methods to quantify patulin-producing molds in food products

机译：开发实时定量PCR方法以定量食品中产生棒曲霉素的霉菌

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摘要

Patulin is a mycotoxin produced by different Penicillium and Aspergillus strains isolated from food products. To improve food safety, the presence of patulin-producing molds in foods should be quantified. In the present work, two real-time (RTi) PCR protocols based on SYBR Green and TaqMan were developed. Thirty four patulin producers and 28 non-producers strains belonging to different species usually reported in food products were used. The patulin production was tested by mycellar electrokinetic capillary electrophoresis (MECE) and high-pressure liquid chromatography-mass spectrometry (HPLC-MS). A primer pair F-idhtrb/ R-idhtrb and the probe IDHprobe were designed from the isoepoxydon dehydrogenase (idh) gene, involved in patulin biosynthesis. The functionality of the developed method was demonstrated by the high linear relationship of the standard curves constructed with the idh gene copy number and Ct values for the different patulin producers tested. The ability to quantify patulin producers of the developed SYBR Green and TaqMan assays in artificially inoculated food samples was successful, with a minimum threshold of 10 conidia g-1 per reaction. The developed methods quantified with high efficiency fungal load in foods. These RTi-PCR protocols, are proposed to be used to quantify patulin-producing molds in food products and to prevent patulin from entering the food chain.
机译：棒曲霉素是一种从食品中分离出的青霉和曲霉菌株产生的霉菌毒素。为了提高食品安全性，应量化食品中出现棒曲霉素的霉菌。在目前的工作中，开发了两种基于SYBR Green和TaqMan的实时（RTi）PCR协议。使用了通常在食品中报告的属于不同物种的34种patulin生产者和28种非生产者菌株。通过Mycellar电动毛细管电泳（MECE）和高压液相色谱-质谱（HPLC-MS）测试棒曲霉素的产生。从异环氧基脱氢酶（idh）基因设计了引物对F-idhtrb / R-idhtrb和探针IDHprobe，参与了棒曲霉素的生物合成。所开发方法的功能性通过用idh基因拷贝数和所测试的不同棒曲蛋白生产商的Ct值构建的标准曲线的高线性关系证明。能够对人工接种的食品样本中已开发的SYBR Green和TaqMan测定法中的棒曲霉素生产者进行定量分析的能力非常成功，每个反应的最低阈值为10分生孢子g-1。所开发的方法对食品中的高效真菌含量进行了定量。建议将这些RTi-PCR方案用于量化食品中产生棒曲霉素的霉菌，并防止棒曲霉素进入食物链。

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  • 来源
    《Food microbiology》 |2011年第6期|p.1190-1199|共10页
  • 作者

    Alicia Rodriguez; M. Isabel Luque; Maria J. Andrade; Mar Rodriguez; Miguel A. Asensio; Juan J. Cordoba;

  • 作者单位

    Higiene y Seguridad Alimentaria, Facultad de Veterinaria, Universidad de Extremadura, Avda. de la Universidad s, 10003-Caceres, Spain;

    Higiene y Seguridad Alimentaria, Facultad de Veterinaria, Universidad de Extremadura, Avda. de la Universidad s, 10003-Caceres, Spain;

    Higiene y Seguridad Alimentaria, Facultad de Veterinaria, Universidad de Extremadura, Avda. de la Universidad s, 10003-Caceres, Spain;

    Higiene y Seguridad Alimentaria, Facultad de Veterinaria, Universidad de Extremadura, Avda. de la Universidad s, 10003-Caceres, Spain;

    Higiene y Seguridad Alimentaria, Facultad de Veterinaria, Universidad de Extremadura, Avda. de la Universidad s, 10003-Caceres, Spain;

    Higiene y Seguridad Alimentaria, Facultad de Veterinaria, Universidad de Extremadura, Avda. de la Universidad s, 10003-Caceres, Spain;

  • 收录信息 美国《科学引文索引》(SCI);美国《化学文摘》(CA);
  • 原文格式 PDF
  • 正文语种 eng
  • 中图分类
  • 关键词

    patulin; quantification; mold; rti-pcr; food; taqman probes; idh;

    机译：棒曲霉素;定量;霉菌;rti-pcr;食品;taqman探针;idh;

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