Brandfass Christoph, Karlovsky Petr
University of Göttingen, Institute of Plant Pathology, Grisebachstrasse 6, 37077 Göttingen, Germany.
BMC Microbiol. 2006 Jan 23;6:4. doi: 10.1186/1471-2180-6-4.
Fusarium head blight (FHB) is a disease of cereal crops, which has a severe impact on wheat and barley production worldwide. Apart from reducing the yield and impairing grain quality, FHB leads to contamination of grain with toxic secondary metabolites (mycotoxins), which pose a health risk to humans and livestock. The Fusarium species primarily involved in FHB are F. graminearum and F. culmorum. A key prerequisite for a reduction in the incidence of FHB is an understanding of its epidemiology.
We describe a duplex-PCR-based method for the simultaneous detection of F. culmorum and F. graminearum in plant material. Species-specific PCR products are identified by melting curve analysis performed in a real-time thermocycler in the presence of the fluorescent dye SYBR Green I. In contrast to multiplex real-time PCR assays, the method does not use doubly labeled hybridization probes.
PCR with product differentiation by melting curve analysis offers a cost-effective means of qualitative analysis for the presence of F. culmorum and F. graminearum in plant material. This method is particularly suitable for epidemiological studies involving a large number of samples.
小麦赤霉病是谷类作物的一种病害，对全球小麦和大麦生产造成严重影响。除了降低产量和损害谷物品质外，小麦赤霉病还会导致谷物被有毒次生代谢产物（霉菌毒素）污染，对人类和牲畜健康构成风险。主要引起小麦赤霉病的镰刀菌属物种是禾谷镰刀菌和燕麦镰刀菌。降低小麦赤霉病发病率的一个关键前提是了解其流行病学。
我们描述了一种基于双重PCR的方法，用于同时检测植物材料中的燕麦镰刀菌和禾谷镰刀菌。通过在存在荧光染料SYBR Green I的实时热循环仪中进行熔解曲线分析来鉴定物种特异性PCR产物。与多重实时PCR检测不同，该方法不使用双标记杂交探针。
通过熔解曲线分析进行产物区分的PCR为植物材料中燕麦镰刀菌和禾谷镰刀菌的存在提供了一种经济高效的定性分析方法。该方法特别适用于涉及大量样品的流行病学研究。
