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首页> 外文会议>International Conference on Environmental, Industrial and Applied Microbiology >Species specific PCR detection protocol for the main mycotoxin-producing Aspergillus species in paprika
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Species specific PCR detection protocol for the main mycotoxin-producing Aspergillus species in paprika

机译：物种特异性PCR检测方案，用于脊髓灰粉中玉米霉菌毒素的曲霉属种类

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Paprika is an important food additive in many countries. Extensive data recently available indicate mycotoxin contamination, being Aspergillus species the most frequently isolated from the paprika samples. The rapid, specific and sensitive methods based on PCR represent useful tools to predict the risk of the most important mycotoxins and to assist strategies to prevent them entering the food chain. In this work, we have developed an integrated protocol to detect the main aflatoxin (A. flavus and A. parasiticus) and ochratoxin A producers (A. niger, A. carbonarius, A. ochraceus, A. westerdijkiae and A. steynii) by specific PCR assays in paprika. We have evaluated two rapid commercial DNA extraction kits and determined the appropriate sample size for detection of the critical toxigenic species (visualization of a specific PCR product on agarose gel). We have analyzed 23 paprika samples using two different sample amounts (0.1 and 1 g) for DNA extraction using two commercial kits. Subsequently, samples were incubated in Sabouraud-Chloramphenicol broth during 0, 1 and 2 days. DNA was tested for specific PCR assays previously developed in our group. Incubation of the samples improved detection of toxigenic Aspergillus species, from 16 % at time 0 to 46% after 2 days of incubation. The highest number of species that could be detected was also observed after two days of incubation, being A. flavus the predominant species. In conclusion, optimal levels of detection were achieved using samples of 1 g of paprika, two days of incubation and with DNeasy Plant Mini Kit extraction kit.
机译：辣椒粉是许多国家的重要粮食添加剂。广泛的数据最近可用表明肌毒素污染，是曲霉属物种，最常与辣椒粉样品分离。基于PCR的快速，特异性和敏感的方法代表了预测最重要的霉菌毒素的风险的有用工具，并协助策略以防止他们进入食物链。在这项工作中，我们开发了一种综合方案，以检测主要的黄曲霉毒素（A.Flavus和A.AxaSiticus）和Ochratoxin作为生产者（A.Niger，A. Carbonarius，A. Ochraceus，A. Westerdijkiae和A. Steynii）辣椒粉中的特异性PCR测定。我们已经评估了两种快速的商业DNA提取试剂盒，并确定了用于检测临界毒性种类的适当样品尺寸（琼脂糖凝胶上的特定PCR产物的可视化）。我们使用两种商业试剂盒分析了使用两种不同的样品量（0.1和1g）进行DNA提取的23种辣椒粉样品。随后，在0,1和2天期间在Sabouraucud-氯霉素液中孵育样品。在我们的组中发育的特定PCR测定测试DNA。孵育样品改善了毒性曲霉物种的检测，在孵育2天后在0至46％的时间内从16％。在孵化两天后，也观察到可以检测到的最多物种数量，是一种毒性物种。总之，使用1g辣椒粉的样品，孵育两天和DNeasy植物迷你试剂盒提取试剂盒，实现了最佳检测水平。

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