PCR-Based Diagnosis of Neoscytalidium dimidiatum Infection Using Internal Transcribed Spacer 1 Region of Ribosomal DNA Primers

Authors

  • Charussri Leeyaphan Laboratory of Space and Environmental Medicine, Graduate School of Medicine, Teikyo University, Tokyo, Japan, and Department of Dermatology, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok 10700, Thailand, and Department of Dermatology, Graduate School of Medicine, Teikyo University, Tokyo, Japan
  • Koichi Makimura Laboratory of Space and Environmental Medicine, Graduate School of Medicine, Teikyo University, Tokyo
  • Chiaki Yamanishi Laboratory of Space and Environmental Medicine, Graduate School of Medicine, Teikyo University, Tokyo
  • Sumanas Bunyaratavej Department of Dermatology, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok 10700
  • Carren Hau Department of Dermatology, Graduate School of Medicine, Teikyo University, Tokyo
  • Yayoi Tada Department of Dermatology, Graduate School of Medicine, Teikyo University, Tokyo
  • Wichit Suthammarak Department of Biochemistry, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok 10700
  • Supannee Kaewsutthi Department of Biochemistry, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok 10700
  • Sutasinee Phaitoonwattanakij Department of Dermatology, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok 10700
  • Shinichi Watanab Department of Dermatology, Graduate School of Medicine, Teikyo University, Tokyo

Keywords:

Neoscytalidium dimidiatum; N. dimidiatum infection; N.dimidiatum-specific primers; PCR-based diagnosis; nondermatophytes

Abstract

Objective: To develop N. dimidiatum-specific single PCR-based identification with DNA sequences of nuclear ribosomal internal transcribed spacer (ITS) 1 region primers to facilitate the rapid and accurate detection of N. dimidiatum.
Methods: N. dimidiatum-specific PCR primers were designed based on the sequence of the internal transcribed spacer 1 region, which is located between 18S and 5.8S nuclear rDNA. Fungal DNA extracted from common causative species for superficial fungal infection including: 2 strains of N. dimidiatum, 9 species of dermatophyte (DMP) and 25 species of non-dermatophyte (NDM) colonies grown on culture plates were used for PCR analysis. Also, 30 clinical specimens collected from 30 patients clinically diagnosed with fungal nail and feet infection who attended Dermatology clinic Siriraj Hospital during October 2015 to November 2015 were used for PCR assay.
Results: Using N. dimidiatum-specific PCR primers, the PCR product was amplified from two standard strains of N. dimidiatum, and there was no amplification from other DMP or NDM species. Regarding sensitivity as lower limit of detection, this PCR method was able to detect 10 pg of N. dimidiatum DNA with ethidium bromide staining and could detect N. dimidiatum in clinical samples.
Conclusion: This newly developed N. dimidiatum-specific PCR identification system is rapid, sensitive, and specific. This diagnostic method will facilitate early and accurate diagnosis and accelerate appropriate treatment in patients with N. dimidiatum infection.

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Published

09-03-2018

How to Cite

Leeyaphan, C., Makimura, K., Yamanishi, C., Bunyaratavej, S., Hau, C., Tada, Y., Suthammarak, W., Kaewsutthi, S., Phaitoonwattanakij, S., & Watanab, S. (2018). PCR-Based Diagnosis of Neoscytalidium dimidiatum Infection Using Internal Transcribed Spacer 1 Region of Ribosomal DNA Primers. Siriraj Medical Journal, 70(1), 28–35. Retrieved from https://he02.tci-thaijo.org/index.php/sirirajmedj/article/view/114900

Issue

Vol. 70 No. 1 (2018): Jan-Feb

Section

Original Article

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