Niessen M L, Vogel R F
Lehrstuhl für Technische Mikrobiologie, Technische Universität München, Freising-Weihenstephan, Germany.
Syst Appl Microbiol. 1998 Dec;21(4):618-31. doi: 10.1016/S0723-2020(98)80075-1.
A PCR based assay (Tox5 PCR) which analyses Fusarium species potentially producing trichothecenes was developed using a pair of primers derived from the DNA-sequence of the trichodiene synthase gene (tri5). The primer pair was tested using DNA isolated from a variety of strains representing 64 species and varieties of Fusarium as well as from other fungi, bacteria and cereals. A 658 bp PCR fragment was specifically amplified with DNA isolated from strains of species belonging to the Fusarium sections Discolor, Sporotrichiella, Arthrosporiella, Gibbosum, and "Dlaminia". PCR products obtained were sequenced. Alignment to tri5 sequences given in the literature revealed a high degree of homology. Results of the PCR developed correlated well with literature data on the trichothecene producing capabilities of the respective species. Potential trichothecene producing fusaria were detected in contaminated cereals and malts using the Tox5 PCR assay. Intensity of the signals produced were well correlated with the concentration of deoxynivalenol (DON) in samples of wheat.
开发了一种基于聚合酶链式反应（PCR）的检测方法（Tox5 PCR），该方法使用从单端孢霉烯合酶基因（tri5）的DNA序列衍生而来的一对引物，分析可能产生单端孢霉烯族毒素的镰刀菌属物种。使用从代表64种镰刀菌属物种和变种以及其他真菌、细菌和谷物的各种菌株中分离的DNA对该引物对进行了测试。用从属于镰刀菌属变色组、分枝孢组、节孢组、瘤座孢组和“Dlaminia”的菌株中分离的DNA特异性扩增出一个658 bp的PCR片段。对获得的PCR产物进行了测序。与文献中给出的tri5序列比对显示出高度同源性。所开发的PCR结果与关于各个物种产生单端孢霉烯族毒素能力的文献数据相关性良好。使用Tox5 PCR检测方法在受污染的谷物和麦芽中检测到了可能产生单端孢霉烯族毒素的镰刀菌。产生的信号强度与小麦样品中脱氧雪腐镰刀菌烯醇（DON）的浓度密切相关。
