Korean J Med Mycol.  1998 Dec;3(2):107-114.

Random Amplified Polymorphic DNA for Classification and Identification of Dermatophytes

• Lee YS
• Yoo JI
• Choi YH
• Joo HY
• Kim BS
• Kim DH

Abstract

BACKGROUND: Dermatophytoses are infections of keratinized tissues, that is, the epidermis, hair and nails, caused by a group of specialized fungi, the dermatophytes. Laboratory diagnoses of dermatophytes such as Tricophyton, Microsporum and Epidermophyton are made by microscopic examination and in vitro culture but they are either time consuming of lacking specificity.
OBJECTIVE
In order to develop and apply more rapid and precise diagnostic tests for fungal pathogens to facilitate the improved identification of dermatophytes, we investigated random amplified polymorphism DNA for classification and identification of dermatophytes.
METHODS
Amplification reactions were performed in volumes of 5011 containing 10mM Tris-HCl(pH 8.3), 50mM KCl, 1.5mM MgCl2, 0.01% (w/v), gelatin, 200mM dNTP mixture, 50pM primer, Taq polymerase (0.025 units/ microliter), DNA 0.001 microgram/microliter. The optimal condition to. PCR was 2 cycles (denaturing 94 degrees C 2min, annealing 33 degrees C 2min, extension 72 degrees C 4min), 40 cycles, and extension (72 degrees C 10min).
RESULTS
RAPD showed interspecies polymorphism in but it had identical patterns in intraspecies.
CONCLUSION
It was confirmed that RAPD PCR analysis with optimal conditions is a fast, economical and reproducible method for identification and classification of dermatophytes isolates.

Keyword

RAPD PCR; Trichopyton; Microsporum; Epidermophyton

MeSH Terms

Arthrodermataceae*
Classification*
Clinical Laboratory Techniques
Diagnostic Tests, Routine
DNA*
Epidermis
Epidermophyton
Fungi
Gelatin
Hair
Magnesium Chloride
Microsporum
Polymerase Chain Reaction
Sensitivity and Specificity
Taq Polymerase
Tinea
DNA
Gelatin
Magnesium Chloride
Taq Polymerase