A Comparison Study between Culture Based Technique and Op-site Non-Culture Based Technique for Identifying Malassezia Yeasts on Normal Skin
2
조회
0
다운로드
1
인용
0
참고문헌
키워드
초록
Background: Culture based technique, a traditional method for extraction of DNA from a cultured colony, was complex in culture conditions and was associated with a lower chance of successful culture. Recently, non-culture based technique, which skipped the culture process and directly extracted fungal DNA and differentiated Malassezia species, has been introduced. Objective: Using 26S rDNA PCR-RFLP, the authors identified Malassezia yeasts and compared the yield of Malassezia DNA by the traditional culture based technique and the non-culture based technique via Op-site adhesive tape. Methods: DNA of Malassezia yeasts were extracted using the culture based technique and the non-culture based technique from normal adults. Comparison was performed in order to clarify the differences between these two techniques. Results: Use of the culture based technique resulted in a culture rate of 57.8% (78 out of 135 samples). On the other hand, using the non-culture based technique, fungal species were identified from all 135samples. Using both techniques, M. globosa was the most identified species. The identification rate of the non-culture based technique was 100%; however, 7 repeats of PCR were required to reach 100%identification. Among samples from five body sites, those from the thigh required 5.5 repeats of PCR. Conclusion: The non-culture based technique was better than the culture based technique. However,due to the low amount of DNA extracts from the body sites with low habitation of Malassezia yeasts,repeated PCR was required for differentiation of Malassezia species.
Abstract
Background: Culture based technique, a traditional method for extraction of DNA from a cultured colony, was complex in culture conditions and was associated with a lower chance of successful culture. Recently, non-culture based technique, which skipped the culture process and directly extracted fungal DNA and differentiated Malassezia species, has been introduced. Objective: Using 26S rDNA PCR-RFLP, the authors identified Malassezia yeasts and compared the yield of Malassezia DNA by the traditional culture based technique and the non-culture based technique via Op-site adhesive tape. Methods: DNA of Malassezia yeasts were extracted using the culture based technique and the non-culture based technique from normal adults. Comparison was performed in order to clarify the differences between these two techniques. Results: Use of the culture based technique resulted in a culture rate of 57.8% (78 out of 135 samples). On the other hand, using the non-culture based technique, fungal species were identified from all 135samples. Using both techniques, M. globosa was the most identified species. The identification rate of the non-culture based technique was 100%; however, 7 repeats of PCR were required to reach 100%identification. Among samples from five body sites, those from the thigh required 5.5 repeats of PCR. Conclusion: The non-culture based technique was better than the culture based technique. However,due to the low amount of DNA extracts from the body sites with low habitation of Malassezia yeasts,repeated PCR was required for differentiation of Malassezia species.
참고문헌 (27)
- Faergemann J, "Age incidence of Pityrosporum orbiculare on human skin", Acta Derm Venereol, 60, pp.531~533 (1980)
- Baillon EH, "Traite de botanique medicale cryptogamique, suivi dutableau de droguier de la Faculte de medecine de Paris" (1889), Doin, Paris
- Ljubojevic S, "The role of Malassezia furfur in dermatology", Clin Dermatol, 20, pp.179~182 (2002)
- Kanda N, "The skin fungus-induced Th1- and Th2- related cytokine, chemokine and prostaglandin E2 production in peripheral blood mononuclear cells from patients with atopic dermatitis and psoriasis vulgaris", Clin Exp Allergy, 32, pp.1243~1250 (2002)
- 임선미, "Molecular Analysis of Malassezia Microflora on the Skin of the Patients with Atopic Dermatitis", Annals of Dermatology, 22(1), pp.41~47 (2010), 대한피부과학회
- 오병호, "Epidemiologic Study of Malassezia Yeasts in Seborrheic Dermatitis Patients by the Analysis of 26S rDNA PCR-RFLP", Annals of Dermatology, 22(2), pp.149~155 (2010), 대한피부과학회
- Leeming JP, "Improved methods for isolation and enumeration of Malassezia furfur from human skin", J Clin Microbiol, 25, pp.2017~2019 (1987)
- Ahn KJ, "Malassezia species cultured from the lesions of pityriasis versicolor", Korean J Dermatol, 35, pp.736~743 (1997)
- Sugita T, "Molecular analysis of Malassezia on the skin of atopic dermatitis patients and healthy subjects", J Clin Microbiol, 39, pp.3486~3490 (2001)
- 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
- Mirhendi H, "A simple PCR-RFLP method for identification and differentiation of 11 Malassezia species", J Microbiol Methods, 61, pp.281~284 (2005)
- Kown KS, "Dermatomycosis, In Korean Dermatological Association, Dermatology. 5th ed", pp.345~364 (2008), Ryomoongak
- Ahn KJ, "Taxonomy of the genus Malassezia", Kor J Med Mycol, 3, pp.81~88 (1998)
- Gueho E, "The genus Malassezia with description of four new species", Antonie van Leeuwenhoek, 69, pp.337~355 (1996)
- Guillot J, "The diversity of Malassezia yeasts confirmed by rRNA sequence and nuclear DNA comparisons", Antonie van Leeuwenhoek, 67, pp.297~314 (1995)
- Sugita T, "New yeast species, Malassezia dermatis, isolated from patients with atopic dermatitis", J Clin Microbiol, 40, pp.1363~1367 (2002)
- Sugita T, "A new yeast, Malassezia yamatoensis, isolated from a patient with seborrheic dermatitis, and its distribution in patients and healthy subjects", Microbiol Immunol, 48, pp.579~583 (2004)
- Sugita T, "Description of a new yeast species, Malassezia japonica, and its detection in patients with atopic dermatitis and healthy subjects", J Clin Microbiol, 41, pp.4695~4699 (2003)
- Hirai A, "Malassezia nana sp. nov., a novel lipid-dependent yeast species isolated from animals", Int J Syst Evol Microbiol, 54, pp.623~627 (2004)
- Cha HG, "The study of incidence and distribution of Pityrospoum on clinically normal skin", Korean J Dermatol, 31, pp.548~558 (1993)
- Kwon HC, "The distribution of Malassezia yeasts on normal human skin by culture study using the scrub-wash technique", Korean J Dermatol, 37, pp.38~45 (1999)
- Kwon HC, "The distribution of Malassezia yeasts on normal human skin by culture study using the swabbing technique", Korean J Dermatol, 37, pp.46~56 (1999)
- Guillot J, "Identification of Malassezia species. A practical approach", J Mycol Med, 6, pp.103~110 (1996)
- 오병호, "Comparison of Nested PCR and RFLP for Identification and Classification of Malassezia Yeasts from Healthy Human Skin", Annals of Dermatology, 21(4), pp.352~357 (2009), 대한피부과학회
- Lee YW, "Quantitative investigation on the distribution of Malassezia species on healthy human skin in Korea", Mycoses, 49, pp.405~410 (2006)
- 장수정, "The Investigation on the Distribution of Malassezia Yeasts on the Normal Korean Skin by 26S rDNA PCR-RFLP", Annals of Dermatology, 21(1), pp.18~26 (2009), 대한피부과학회