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Random Amplified Polymorphic DNA for Classification and Identification of Dermatophytes

Yeong Seon Lee,Jae Il Yoo,Yeon Hwa Choi,Hyung Yeul Joo,Bong Su Kim,Dong Han Kim
Epub 2016 February 24

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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 50 μl containing 10 mM Tris-HCl (pH 8.3), 50 mM KCl, 1.5 mM MgCl2, 0.01% (w/v), gelatin, 200 mM dNTP mixture, 50 pM primer, Taq polymerase (0.025 units/μl), DNA 0.001 μg/μl. The optimal condition to. PCR was 2 cycles (denaturing 94℃ 2 min, annealing 33℃ 2 min, extension 72℃ 4 min), 40 cycles, and extension (72℃ 10 min).


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.



Keywords


Epidermophyton Microsporum RAPD PCR Trichopyton




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