A new molecular marker for species-specific identification of Microsporum canis

Abstract

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.

Remarkable Phenotypic Virulence Factors of Microsporum canis and Their Associated Genes: A Systematic Review

“…M. canis is a fungus of medical importance to both humans and pets. Due to its characteristics, M. canis is one of the main etiological agents of tineas and dermatophytoses worldwide [ 2 , 3 , 7 , 9 , 23 , 28 , 30 , 32 , 36 , 43 , 44 , 45 ]. The severity of the infection produced in some susceptible hosts and the difficulties that arise in establishing treatments or prevention strategies are the main causes of its dissemination [ 5 , 7 , 11 , 12 , 13 ].…”

“…Among the most common dermatophytes are the three species of the genus Microsporum : M. audouinii and M. ferrugineum , both anthropophilic, and M. canis , which is considered anthropo-zoophilic; all of them are associated with dermatophytosis in domestic animals and tinea capitis, tinea corporis and occasionally onychomycosis in humans [ 2 , 6 , 7 , 8 , 9 , 10 ]. In particular, M. canis is considered to be of medical importance because it is capable of infecting pediatric or immunocompromised patients.…”

Molecular identification of Microsporum canis isolates and detection of subtilisin genes from dermatophytosis

“…Since dermatophytes is a major health problem in Egypt, a rapid and specific identification for them should be available [18]. In our study we compared the detection of M.canis using the [19]. This is due to, the nucleotide sequence of the velB gene contains a unique microsatellite motif (CAGCAC)8 for identification of M. canis.…”

“…This is due to, the nucleotide sequence of the velB gene contains a unique microsatellite motif (CAGCAC)8 for identification of M. canis. These species-specific primers can distinguish M. canis from the other phylogenetically closely related species that can cause similar clinical manifestations [19].…”

First study on molecular epidemiology of dermatophytosis in cats, dogs, and their companions in the Kurdistan region of Iraq

“…DNA was extracted from positive dermatophytes’ cultures using a fungal DNA preparation kit (Jena Bioscience, Germany). Two sets of primers were used, Mc-VelB-F (5′-CTTCCCCACCCGCAACATC-3′) and Mc-VelB-R (5′-TGTGGCTGCACCTGAGAGT GG-3′), for M. canis [ 7 ]. Whereas, CHS1 gene primers, the forward CHS1 1S (5′-CAT CGA GTA CAT GTG CTC GC-3′) and the reverse CHS1 1R (5′-CTC GAG GTC AAA AGC ACG CC-3′), were used for Trichophyton mentagrophytes [ 8 ].…”