Abstract
Dermatophytes being animal and human pathogenic fungi infect some human at one point or the other in their lifetime. For effective control of dermatophytes, accurate identification of the specific species/strain involved must be known. Stocks from pathogenic fungi isolated from infected areas on different patients, around Lagos-Nigeria were analysed using molecular methods (DNA extraction, PCR-RFLP and DNA sequencing). Four DNA extraction protocols were employed in the identification of the fungal isolates. Sixteen different fungal isolates were identified, and based on the molecular data these were classified into six species of dermatophytes belonging to the genera Microsporum, Trichophyton and Epidermaphyton, two species of systemic mycoses fungi and eight opportunistic human pathogenic fungi. The results also revealed that CTAB protocol, Modified CTAB protocol and the GNOME kit used in this work were only able to extract non-dermatophytes DNA. Only the Zymo DNA kit was able to isolate dermatophytes DNA. DNA extraction which is the first step in all molecular studies showed that one DNA extraction method might not be able to extract all fungal DNA for proper identification, diagnosis and treatment of fungal infections. Keywords: Dermatophytes, DNA extraction, Identification, Protocols.
Highlights
Dermatomycoses is contagious and represents a significant health problem in Nigeria
decanted gently and the pellet (DNA) extraction using Cetyltrimethyl ammonium bromide (CTAB) protocol: Ten millilitres (10 ml) of sterilized distilled water was added to fresh pure culture of each isolate to be extracted, a suspension was made by gently probing the colony with the tip of a sterilised Pasteur pipette
Plate 1a shows the electrophorogram of fungal DNA extracted with the CTAB protocol, this protocol extracted DNA of all the fungal isolates and showed good bands on the electrophorogram expect for the dermatophytes isolates (Table 1)
Summary
Lagos metropolis has a higher incidence of dermatomycoses due to crowded living conditions (Adetosoye, 1977; Ogbonna et al, 1985; Anosike et al, 2005). Incidences of the prevalence is particular to Lagos-Nigeria but is observed in many countries and crowded places in the world. The current diagnosis of dermatomycoses in most mycological laboratories inclusive of Lagos state, Nigeria is based upon conventional methods which is not species-specific. This does not give the accurate diagnostic results as the etiological patterns of most pathogenic organisms are changing just as there is change in climate. This study aims at isolating and identifying this group of human pathogenic fungi responsible for dermatomycoses using molecular methods
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