Identification and characterization of dermatophyte species and strains with PCR amplification.

Guofang Liu,Chenghua He,Haibin Zhang

doi:10.3892/etm.2014.1785

Guofang Liu, Chenghua He + Show 1 more

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https://doi.org/10.3892/etm.2014.1785

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Abstract

The aim of the present study was to use two polymerase chain reaction (PCR) methods, with (GACA)4 and non-transcribed spacer (NTS) as primers, to identify and characterize dermatophyte isolates from dogs and cats to a species and strain level. A total of 45 isolates from nine dermatophyte species were collected from pet dogs and cats and subjected to PCR amplification with the microsatellite primer (GACA)4. Dermatophyte strains of three of the same species collected from four cities were subjected to PCR amplification with the NTS primer set. These two PCR methods were applied to identify and characterize the dermatophyte isolates to a species and strain level. Regional differences among the strain specificities were also examined. The results from PCR with (GACA)4 demonstrated that strains from the same species produced similar PCR product band patterns. In addition, these patterns differed among species, indicating that (GACA)4 primer-based PCR was able to distinguish between the various dermatophyte species. By contrast, dermatophyte isolates and/or strains within the same species revealed various band patterns with NTS-based PCR. In addition, the results indicated that regional differences contributed to the variations in PCR product band patterns. Therefore, the results of the present study indicate that the NTS-based PCR method is efficient in distinguishing dermatophytes to the strain level, while a combination of (GACA)4 and NTS primer-based PCR methods is able to clarify dermatophyte isolates to a species and strain level. The present study provides information concerning the identification of pathogenic fungi and the epidemiological characteristics of fungal skin diseases.

Highlights

Fungal infections affect superficial keratinized tissues, including the skin, hair and nails, in humans and animals resulting in difficult‐to‐treat dermatosis
Based on clinical phenotypic analysis, strains from the same species produced similar patterns, but these patterns changed from species to species (Fig. 1). These results indicate that (GACA)4 primer‐based polymerase chain reaction (PCR) is able to distinguish between various dermatophyte species, which may be useful for species identification
The non‐transcribed spacer (NTS)‐2 amplification products for the various strains exhibited the same profile, consisting of two clearly distinguishable bands of 800 and 650 bp (Fig. 5B). These results indicate that dermatophyte isolates and/or strains within the same species exhibit various band patterns with NTS‐based PCR, indicating that this method may be a useful tool to identify dermatophytes to the strain level

Summary

Introduction

Fungal infections affect superficial keratinized tissues, including the skin, hair and nails, in humans and animals resulting in difficult‐to‐treat dermatosis. Fungi derived from pet dogs and cats, including dermatophytes (Deuteromycotina, Hyphomycetes, Hyphomycetales, Moniliaceae, Trichophyton, Microsporum and Epidermophyton), Malassezia, Saccharomycetes (mainly Candida) and non‐dermatophyte molds (Scopulariopsis, Aspergillus and Fusarium), are able to infect human skin [1]. The simple repetitive oligonucleotide, (GACA), is a highly variable microsatellite that has been used as a PCR primer for the efficient identification of skin tinea infections and pathogenic Candida species [4]. PCR using (GACA) has been used for the classification and identification of human pathogenic fungi [5]. Microsatellite (GACA) and non‐transcribed spacer (NTS) primers were used to perform PCR amplification with the aim of identifying and characterizing dermatophyte isolates from dogs and cats to a species and strain level

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