Evaluation of growth conditions and DNA extraction techniques used in the molecular analysis of dermatophytes

Abstract

Most mycoses are caused by fungi that reside in nature. The severity of the infection depends on the pathogenic attributes, socioeconomic factors and local environmental conditions. Recent diagnosis increasingly relies on not only the clinical features. Molecular identifications have determined the need for a rapid and easy method of extracting DNA. Usually two factors have to be considered: maximize the DNA yield and ensure that the extracted DNA is susceptible to enzymatic reactions. These data suggest that phenol-chloroform methods and a 7-day culture period may be useful for validation and constitute the first step of molecular diagnosis of dermatophytes.

Infection of Trichophyton verrucosum in cattle breeders, Poland: A 40‐year retrospective study on the genomic variability of strains

“…In the literature, there are no reports of the use of these techniques for dermatophyte identification in a wide range by many laboratories. In their investigations, Faggi et al, Brillowska‐Dąbrowska et al, Shehata et al , and Gnat et al show that this method can be a useful tool in molecular identification of tested isolates to the species level in full agreement with both culture‐based and ITS‐based methods. Our study also revealed that the PCR‐based band profiles obtained by MP‐PCR and PCR with primers (GTG) 5 and (GACA) 4 were reliable at the same level as ITS analyses, and were additionally easier to do.…”

“…DNA was isolated from the dermatophytes with the phenol‐chloroform method as described by Gnat et al . Extraction controls were generated by subjecting distilled water to the same procedure.…”

“…However, anthropophilic species have been found to cause infections in animals . In the last several years, infections caused by dermatophytes have increased dramatically, and it is currently estimated that superficial mycoses affect more than 20%‐25% of the world's population . They have also recently gained prominence due to their rising incidence in patients with immunocompromised status such as cancer, diabetes mellitus, AIDS and organ transplantation .…”

“…Furthermore, dermatophytes are also repeatedly cited among the most frequent causes of dermatological problems in animals, especially domestic . An active infection in animals is not common, but many animals can be dermatophyte carriers . Factors that can predispose animals to an infection are above all stress, diet, poor environmental conditions (heat, humidity and overcrowding), age and genetic predispositions .…”

“…An active infection in animals is not common, but many animals can be dermatophyte carriers . Factors that can predispose animals to an infection are above all stress, diet, poor environmental conditions (heat, humidity and overcrowding), age and genetic predispositions . Humans are usually infected from animals mostly through direct contact or indirectly via fungus‐bearing hair and scales from infected animals …”