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A molecular diagnostic assay for the detection and identification of wood decay fungi of conifers

机译：针叶林木材腐烂真菌检测与鉴定的分子诊断方法

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Ten taxon-specific primers were designed to amplify the Internal Transcribed Spacer of the rRNA operon of several important decay fungi of coniferous wood, including Armillaria spp., Echinodontium spp., Fomitopsis pinicola, Fuscoporia torulosa, Heterobasidion annosum sensu lato (s.l.), Onnia spp., Phaeolus schweinitzii, Phellinus weirii s.l., Pholiota spp. and Porodaedalea spp. Primers designed in this study and in a previous one for the identification of Laetiporus sulphureus and Stereum spp. were combined in two multiplex PCRs, which were tested for efficiency and specificity, and detected at least 1pg of fungal target DNA. Target DNA at concentrations of 10(-1)pg or lower can be detected with this assay using SYBR (R) Green Real-Time PCR. Validation assays performed on 129 naturally infected wood samples or fruiting bodies confirmed the reliability of the multiplex PCR-based diagnostic method. This method represents a simple and rapid diagnostic tool for the detection of a number of destructive wood decay fungi of conifer wood.
机译：设计了十种分类群特异性引物，以扩增针叶木材的几种重要衰变真菌rRNA操纵子的内部转录间隔子，这些真菌包括：蜜环菌属，棘皮虫属，松叶拟南芥，扶桑孢子虫，无名氏异位藻（Son），Onnia ，Phaeolus schweinitzii，Phellinus weirii sl，Pholiota spp。和Porodaedalea spp。在本研究和上一个研究中设计的引物用于鉴定Laetiporus sulphureus和Stereum spp。将其组合在两个多重PCR中，测试其效率和特异性，并检测至少1pg的真菌靶DNA。可以使用SYBR Green Real-Time PCR通过此测定法检测浓度为10（-1）pg或更低的目标DNA。对129种自然感染的木材样品或子实体进行的验证分析证实了基于多重PCR的诊断方法的可靠性。该方法代表了一种简单而快速的诊断工具，用于检测针叶树木材的许多破坏性木材腐烂真菌。

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