RAPD PCR분석에 의한 국내 피부 사상균속 분류 및 동정 = Random Amplified Polymorphic DNA for Classification and Identification of Dermatophytes

다국어 초록 (Multilingual Abstract)

Backgound: 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 Trichophyton, Microsporum and Epidermophyton are made by microscopic examination and in vitro culture but they are either time consuming or 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㎕ containing 10mM Tris-HCl (pH 8.3), 50mM KCI, 1.5mM MgCl_2 0.01% (w/v), gelatin, 200mM dNTP mixture, 50pM primer, raq polymerase (0.025units/㎕), DNA 0.O01㎕. The optimal condition for PCR was 2 cycles (denaturing 94℃ 2min, annealing 33℃ 2min, extention 72℃ 4miu), 40 cycles, and extention (72℃ 10min).
Results: RAPD showed interspecies polymorphism in T. rubum, T. mentagrophytes, T. tonsurans, T. violaceum, M. ferrugineum, M. canis, M. gypseum, M. audouinii and E. floccosum, 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.

목차 (Table of Contents)

• 서론
• 재료 및 방법
• 1. 실험 재료
• 2. 실험 방법
• 결 과
• 1. PCR 최적 조건 확립
• 2. 표준 균주에 대한 RAPD 분석
• 3. 임상 분리주에 대한 RAPD 분석
• 고 찰
• 결 론