Department of Animal Production, College of Agriculture, Al-Qasim Green University, Al-Qasim 51013, Babil, Iraq.
Department of Plant Protection, College of Agriculture, University of Kufa, Al-Kufa, Najaf 54001, Iraq.
Microb Pathog. 2018 Mar;116:351-355. doi: 10.1016/j.micpath.2018.01.049. Epub 2018 Feb 7.
Aspergillus flavus is among the most devastating opportunistic pathogens of several food crops including rice, due to its high production of carcinogenic aflatoxins. The presence of these organisms in economically important rice strip farming is a serious food safety concern. Several polymerase chain reaction (PCR) primers have been designed to detect this species; however, a comparative assessment of their accuracy has not been conducted. This study aims to identify the optimal diagnostic PCR primers for the identification of A. flavus, among widely available primers. We isolated 122 A. flavus native isolates from randomly collected rice strips (N = 300). We identified 109 isolates to the genus level using universal fungal PCR primer pairs. Nine pairs of primers were examined for their PCR diagnostic specificity on the 109 isolates. FLA PCR was found to be the optimal PCR primer pair for specific identification of the native isolates, over aflP(1), aflM, aflA, aflD, aflP(3), aflP(2), and aflR. The PEP primer pair was found to be the most unsuitable for A. flavus identification. In conclusion, the present study indicates the powerful specificity of the FLA PCR primer over other commonly available diagnostic primers for accurate, rapid, and large-scale identification of A. flavus native isolates. This study provides the first simple, practical comparative guide to PCR-based screening of A. flavus infection in rice strips.
黄曲霉是包括水稻在内的几种粮食作物中最具破坏性的机会致病菌之一，因为它能产生致癌的黄曲霉毒素。这些生物在经济上重要的水稻条播种植中存在，是一个严重的食品安全问题。已经设计了几种聚合酶链反应（PCR）引物来检测这种物种；然而，尚未对其准确性进行比较评估。本研究旨在确定最适合用于鉴定黄曲霉的诊断性 PCR 引物，这些引物在广泛可用的引物中。我们从随机采集的水稻条带（N=300）中分离出 122 株黄曲霉本地分离株。我们使用通用真菌 PCR 引物对将 109 株分离株鉴定到属水平。检查了 9 对引物对 109 株分离株的 PCR 诊断特异性。发现 FLA PCR 是鉴定本地分离株的最佳 PCR 引物对，优于 aflP(1)、aflM、aflA、aflD、aflP(3)、aflP(2)和 aflR。PEP 引物对被发现最不适合鉴定黄曲霉。总之，本研究表明，FLA PCR 引物的特异性强于其他常用的诊断性引物，可用于准确、快速和大规模鉴定黄曲霉本地分离株。本研究为基于 PCR 的水稻条带黄曲霉感染筛查提供了第一个简单实用的比较指南。
