Simultaneous detection of Fusarium culmorum and F. graminearum in plant material by duplex PCR with melting curve analysis
Simultaneous detection of Fusarium culmorum and F. graminearum in plant material by duplex PCR with melting curve analysis
复制标题
通过双重 PCR 和熔解曲线分析同时检测植物材料中的镰刀菌和禾谷镰刀菌
作者：
C. Brandfass;Petr Karlovsky
镰刀菌头疫病（fusarium head blight， FHB）是一种严重影响全球小麦和大麦生产的谷类作物病害。除了降低产量和损害粮食质量外，食品血红蛋白还会导致有毒次生代谢物（真菌毒素）污染粮食，从而对人类和牲畜构成健康风险。赤霉病主要涉及的镰刀菌种类是F. graminearum和F. culmorum。减少FHB发病率的一个关键先决条件是了解其流行病学。结果建立了一种基于双pcr的同时检测植物材料中镰刀菌和谷粒镰刀菌的方法。物种特异性PCR产物通过实时热循环仪在荧光染料SYBR Green i的存在下进行熔化曲线分析来鉴定。与多重实时PCR分析相比，该方法不使用双重标记杂交探针。结论采用熔点曲线分析进行产物分化的pcr方法是一种有效的定性分析植物材料中镰刀菌和谷草镰刀菌的方法。这种方法特别适用于涉及大量样本的流行病学研究。
BackgroundFusarium 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.ResultsWe 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.ConclusionPCR 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.
影响因子：
2.7
作者：
Stewart Cn;L. Via
通讯作者：
Stewart Cn;L. Via