Species identification of dermatophytes by conventional mycological methods based on macro- and microscopy analysis is time-consuming and has a lot of limitations such as slow fungal growth or low specificity. Thus, there is a need for the development of molecular methods that would provide reliable and prompt identification of this group of medically important fungi. The are many reports in the literature concerning PCR identification of dermatophyte species, but still, there are not many PCR assays for the separate detection of members of the genera Microsporum, especially Microsporum canis (zoophilic species) and Microsporum audouinii (anthropophilic species). The correct distinction of these species is important to determine the source of infection to implement the appropriate action to eliminate the path of infection transmission. In this paper, we present such a PCR-based method targeting velB gene that uses a set of two primers—Mc-VelB-F (5′-CTTCCCCACCCGCAACATC-3′) and Mc-VelB-R (5′-TGTGGCTGCACCTGAGAGTGG-3′). The amplified fragment is specific due to the presence of (CAGCAC)8 microsatellite sequence only in the velB gene of M. canis. DNA from 153 fungal samples was used in PCR assay followed by electrophoretic analysis. The specificity of the designed set of primers was also confirmed using the online BLAST-Primer tool. The positive results were observed only in the case of M. canis isolates, and no positive results were obtained neither for other dermatophytes and non-dermatophyte fungi nor for other Eukaryotes, including the human genome sequence, as well as the representatives of bacterial and viral taxa. The developed PCR assay using the proposed Mc-VelB-F and Mc-velB-R primers can be included in the algorithm of M. canis detection in animals and humans.

中文翻译：

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犬小孢子菌种特异性鉴定的新分子标记

通过基于宏观和显微镜分析的传统真菌学方法对皮肤癣菌进行物种鉴定非常耗时，并且具有许多局限性，例如真菌生长缓慢或特异性低。因此，需要开发能够可靠且迅速地鉴定这组医学上重要的真菌的分子方法。文献中关于皮肤癣菌物种的 PCR 鉴定有很多报道，但仍然没有很多 PCR 检测用于分别检测小孢子菌属的成员，尤其是犬小孢子菌（动物性物种）和小孢子菌属（嗜人性物种）。正确区分这些物种对于确定感染源以实施适当的行动以消除感染传播途径很重要。在本文中，我们提出了一种基于 PCR 的靶向 velB 基因的方法，该方法使用一组两个引物——Mc-VelB-F (5'-CTTCCCCCACCCGCAACATC-3') 和 Mc-VelB-R (5'-TGGTGGCTGCACCTGAGAGTGG-3 ')。由于 (CAGCAC)8 微卫星序列仅在犬支原体的 velB 基因中存在，因此扩增片段具有特异性。来自 153 个真菌样品的 DNA 用于 PCR 分析，然后进行电泳分析。还使用在线 BLAST-Primer 工具确认了设计的引物组的特异性。仅在犬分枝杆菌分离株中观察到阳性结果，对于其他皮肤癣菌和非皮肤癣菌真菌以及其他真核生物，包括人类基因组序列，以及细菌和病毒的代表均未获得阳性结果。分类群。