Abstract

Background: In Korea, Trichophyton (T.) rubrum is occupied more than 80% of causative fungi of dermatophytosis. Strain differentiation of T. rubrum is essential for epidermiologic study.

Objective: The aim of this study is to develope the primers for amplification of rDNA intergenic spacer(IGS) to detect polymorphism of T. rubrum.

Methods: The primers were designed from 25S and 18S of rDNA, and were applied to 20 strains of T. rubrum, which included 1 standard strain and 19 clinical isolates.

Results: Primers for amplification of polymorphic rDNA IGS were designed from the 3’end of the 25S (primer ANID25-3, 5’-GACAGGTTAGTTTTACCCTACTGA-3’) and the 5’end of the 18S (TR18-2R, 5’-ATCTAATAAATACACCCCTTCCGA-3’). PCR condition was adjusted for detecting polymorphism. Best results were obtained at 55℃ for annealing temperature and 3 minutes for extension time. Eight bands sized as 1.1, 2.4, 2.7, 2.9, 3.2, 3.8, 5 kb were amplified with PCR using the primers. With 4 bands sized 2.7, 2.9, 3.2 and 3.8 kb, 20 strains of T. rubrum could be grouped into 6 subtypes.

Conclusion: The PCR fingerprinting with the primers for rDNA IGS was able to differentiated strains of T. rubrum, and it can be applied in clinical and epidemiologic studies. The primer could be applied to other fungi with unknown sequences.

Keywords

Trichophyton rubrum rDNA intergenic spacer Polymorphism

KJMM 2004 December;9(4):206-213(8). Epub 2016 February 20
Copyright © 2004 by Korean Journal of Medical Mycology

Language

Korean/English

Author

Jong Soo Choi; Department of Dermatology, College of Medicine, Yeungnam University, Daegu, Korea
Dong Hoon Shin; Department of Dermatology, College of Medicine, Yeungnam University, Daegu, Korea
Jun Je Seung; Dr. Seung’s Skin Clinic, Gumi, Korea
Ki Hong Kim; Department of Dermatology, College of Medicine, Yeungnam University, Daegu, Korea

Corresponding

Jong Soo Choi, Department of Dermatology, College of Medicine, Yeungnam University, Daegu, Korea. Tel: (053) 620-3160, Fax: (053) 622-2216, e-mail: jschoi@med.yu.ac.kr

Publication history

Acknowledgements

This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

Jong Soo Choi

Department of Dermatology, College of Medicine, Yeungnam University, Daegu, Korea

Dong Hoon Shin

Department of Dermatology, College of Medicine, Yeungnam University, Daegu, Korea

Jun Je Seung

Dr. Seung’s Skin Clinic, Gumi, Korea

Ki Hong Kim

Department of Dermatology, College of Medicine, Yeungnam University, Daegu, Korea

Since epub date 2016 February 20