Malassezia 효모균 동정을 위한 Multiplex PCR 기법 개발과 평가
The Development and Evaluation of Multiplex PCR Technique for Identification of Malassezia Yeast
2
조회
0
다운로드
2
인용
0
참고문헌
키워드
초록
Background: Malassezia yeasts as major pathogenic fungi causes the common skin diseases including dandruff, psoriasis, seborrheic dermatitis and atopic dermatitis etc. various molecular techniques were developed to identify and classify the Malassezia species until now. But, these methods were discovered the problems. So, the development of the better molecular methods required to identify and classify of Malasseiza species. Objective: We sought to develop of molecular techniques to identify and classify of six Malassezia species (M. restricta, M. globosa, M. furfur, M. slooffiae, M. sympodialis, M. obtusa). Methods: We designed primers about ITS1 (Internal transcribed space 1) region that were wellknown region useful to identify of Malassezia species. Because, ITS1 region that is located between 18S and 5.8S rDNA of ribosomal DNA was comparatively mutated quickly. The mono PCR using ITS1 primers was performed to confirm the specificity of ITS1 primers with six Malassezia standard strains. Then, Malassezia Multiplex detection kit was developed on the basis of technique using ITS1regions. Malassezia Multiplex detection kit was used to perform multiplex PCR with six Malassezia standard strains and clinical isolates. Results: The results of mono PCR using ITS1 primers about six Malassezia standard strains was detected each Malassezia standard strains. Also, the multiplex PCR using developed Malassezia Multiplex detection kit was confirmed to classify about six Malassezia standard strains and clinical isolates. Conclusion: In this study, we verified that six Malassezia yeasts was classified using Malassezia Multiplex detection kit from Malasszia standard strains and clinical isolates. And we anticipate that Malassezia Multiplex detection kit is able to do accurate diagnosis about six Malassezia yeasts (M.restricta, M. globosa, M. furfur, M. slooffiae, M. sympodialis, M. obtusa).
Abstract
Background: Malassezia yeasts as major pathogenic fungi causes the common skin diseases including dandruff, psoriasis, seborrheic dermatitis and atopic dermatitis etc. various molecular techniques were developed to identify and classify the Malassezia species until now. But, these methods were discovered the problems. So, the development of the better molecular methods required to identify and classify of Malasseiza species. Objective: We sought to develop of molecular techniques to identify and classify of six Malassezia species (M. restricta, M. globosa, M. furfur, M. slooffiae, M. sympodialis, M. obtusa). Methods: We designed primers about ITS1 (Internal transcribed space 1) region that were wellknown region useful to identify of Malassezia species. Because, ITS1 region that is located between 18S and 5.8S rDNA of ribosomal DNA was comparatively mutated quickly. The mono PCR using ITS1 primers was performed to confirm the specificity of ITS1 primers with six Malassezia standard strains. Then, Malassezia Multiplex detection kit was developed on the basis of technique using ITS1regions. Malassezia Multiplex detection kit was used to perform multiplex PCR with six Malassezia standard strains and clinical isolates. Results: The results of mono PCR using ITS1 primers about six Malassezia standard strains was detected each Malassezia standard strains. Also, the multiplex PCR using developed Malassezia Multiplex detection kit was confirmed to classify about six Malassezia standard strains and clinical isolates. Conclusion: In this study, we verified that six Malassezia yeasts was classified using Malassezia Multiplex detection kit from Malasszia standard strains and clinical isolates. And we anticipate that Malassezia Multiplex detection kit is able to do accurate diagnosis about six Malassezia yeasts (M.restricta, M. globosa, M. furfur, M. slooffiae, M. sympodialis, M. obtusa).
참고문헌 (15)
- 이미경, "다중 중합효소연쇄반응을 이용한 Candida 균종동정", 대한임상미생물학회지, 9(2), pp.119~124 (2006), 대한임상미생물학회
- Koichi M, "species identification and strain typing of Malassezia species stock strains and clinical isolates based on the DNA sequences of nuclear ribosomal internal transcribed spacer 1 regions", J Med Microbiol, 49, pp.29~35 (2000)
- Aditya K, "Molecular differentiation of seven Malassezia species", J Clin Microbiol, 38(5), pp.1869~1875 (2000)
- 김상민, "말라쎄지아 효모균의 분자생물학적 분석으로서집락 중합효소연쇄반응의 적용", 대한의진균학회지, 12(4), pp.180~188 (2007), 대한의진균학회1
- 이양원, "Malassezia 효모균 동정에 26S rDNA PCR-RFLP기법의 적용", 대한의진균학회지, 11(3), pp.141~153 (2006), 대한의진균학회1
- Christina M, "Fast, noninvasive method for molecular detection and differentiation of Malassezia yeast species on human skin and application of the method to Dandruff microbiology", J Clin Microbiol, 40(9), pp.3350~3357 (2002)
- 임세웅, "환자 피부 비늘 및 임상주에서 이중 중합효소 연쇄반응(Nested PCR)을 이용한 Malassezia 균종의 동정", 대한피부과학회지, 46(4), pp.446~452 (2008), 대한피부과학회
- 송영찬, "Pyrosequencing 방법을 이용한 Malassezia 효모균의분자생물학적 동정", 대한의진균학회지, 12(4), pp.189~197 (2007), 대한의진균학회1
- 김범준, "Optimal Culture Condition for Antifungal Susceptibility Tests of Malassezia globosa", 대한의진균학회지, 14(4), pp.182~189 (2009), 대한의진균학회1
- Takamasa K, "Revised culture-based system for identification of Malassezia species", J Clin Microbiol, 45(11), pp.3737~3742 (2007)
- Gaitanis G, "Identification of Malassezia species from patient skin scales by PCRRFLP", Clin Microbiol Infect, 8, pp.162~173 (2002)
- George G, "The range of molecular methods for typing Malassezia", Current Opinion in Infectious Diseases, 22, pp.119~125 (2009)
- Bart T, "Identification and typing of Malassezia yeasts using amplified fragment length polymorphism (AFLPTM), random amplified polymorphic DNA (RAPD) and denaturing gradient gel electrophoresis (DGGE)", FEMS Yeast Research, 1(2), pp.79~86 (2001)
- AJ Kindo, "Identification of Malassezia species", Indian Journal of Medical Microbiology, 22(3), pp.179~181 (2004)
- Guého E, "The genus Malassezia with description of J four new species", Antonie van Leeuwenhoek, 69, pp.337~355 (1996)