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Direct Species Identification of Common Pathogenic Dermatophyte Fungi in Clinical Specimens by Semi-nested PCR and Restriction Fragment Length Polymorphism

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Abstract

Objective

To seek a rapid and reliable molecular biology method to identify the common pathogenic dermatophyte fungi from clinical samples.

Method

The genome DNA was extracted from cultured strains of seven common dermatophyte fungi species and part of each positive clinical specimen by microscopy. Intergenic spacer regions of ribosomal DNA (ITS) were amplified by semi-nested PCR (snPCR) with three universal primers (NS5, ITS1, and ITS4) for fungi. The amplified products were digested with two restriction endonucleases (BciT130 I, Dde I), the Restriction Fragment Length Polymorphism(RFLP). The rest of each clinical specimen was cultured in Sabouraud’s Agar medium. Then the results of RFLP were compared with the traditional culture results.

Results

The digestion of seven common dermatophyte fungi produced seven different restriction profiles. Restriction profiles of 17 clinical specimens matched, respectively, to that of the cultured strains, and 14 profiles of the 17 ones matched the culture result completely. The coincidence was 100.0%.

Conclusions

snPCR-RFLP analysis of intergenic spacer regions of ribosomal DNA is a valuable method of exactness and clarity for species identification of common dermatophyte fungi from clinical specimens.

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Authors and Affiliations

  1. Department of Dermatology, The 1st Affiliated Hospital of Dalian Medical University, 116011, Dalian, China

    Guoling Yang & Mingli Zhang

  2. Department of Bioengineering, Dalian University of Technology, 116024, Dalian, China

    Wenli Li & Lijia An

Authors
  1. Guoling Yang
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  2. Mingli Zhang
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  3. Wenli Li
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  4. Lijia An
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Correspondence to Guoling Yang.

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Yang, G., Zhang, M., Li, W. et al. Direct Species Identification of Common Pathogenic Dermatophyte Fungi in Clinical Specimens by Semi-nested PCR and Restriction Fragment Length Polymorphism. Mycopathologia 166, 203–208 (2008). https://doi.org/10.1007/s11046-008-9130-3

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  • DOI: https://doi.org/10.1007/s11046-008-9130-3

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