Identification of Dermatophytes by Polymerase Chain Reaction-Restriction Fragment Length Polymorphism Analysis of Metalloproteinase-1
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Background: Transgenic research on metalloproteinase-1 isan emerging field in the area of plant molecular biology. Thenew method reported here can similarly be applied in fungalmolecular biology to identify different dermatophytes. Ourmethod is more accurate than traditional methods such asmolecular analyses. Objective: To identify Trichophytonrubrum, T. mentagrophytes var. mentagrophytes, T. tonsurans,T. mentagrophytes var. interdigitale, Microsporumcanis and M. gypseum, by using the restriction fragmentlength polymorphism (RFLP) analysis and polymerase chainreaction (PCR) to detect polymorphisms in the metalloproteinase-1 gene (MEP1). Methods: From each fungal strain, weisolated genomic DNA and performed PCR to amplify theregion coding for metalloproteinase-1. Primers for the metalloproteinase-1 gene were designed based on the sequencein NCBI GenBank. Subsequently, we purified the amplifiedPCR product and performed RFLP analysis. After restrictionenzyme digestion, BsrDI (NEB, England), the samples weresubjected to electrophoresis. Four different patterns of DNAfragments were observed among 6 fungal species. Results:The DNA fragments for T. mentagrophytes var. mentagrophytes,T. mentagrophytes var. interdigitale and T. tonsuransshowed similar patterns on electrophoresis and werenot distinguishable, whereas T. rubrum, M. canis, and M. gypseum showed different patterns. Conclusion: To ourknowledge, it is the first study to introduce the analysis of thenucleotide sequence of metalloproteinase-1 enzyme tostudy differentiation in dermatophytes. Based on our results,more accurate differentiation and subtyping of T. rubrum andT. mentagrophytes var. interdigitale might be possible. Thismight contribute to better understanding of the epidemiologyand pathogenesis of dermatophyte.
Abstract
Background: Transgenic research on metalloproteinase-1 isan emerging field in the area of plant molecular biology. Thenew method reported here can similarly be applied in fungalmolecular biology to identify different dermatophytes. Ourmethod is more accurate than traditional methods such asmolecular analyses. Objective: To identify Trichophytonrubrum, T. mentagrophytes var. mentagrophytes, T. tonsurans,T. mentagrophytes var. interdigitale, Microsporumcanis and M. gypseum, by using the restriction fragmentlength polymorphism (RFLP) analysis and polymerase chainreaction (PCR) to detect polymorphisms in the metalloproteinase-1 gene (MEP1). Methods: From each fungal strain, weisolated genomic DNA and performed PCR to amplify theregion coding for metalloproteinase-1. Primers for the metalloproteinase-1 gene were designed based on the sequencein NCBI GenBank. Subsequently, we purified the amplifiedPCR product and performed RFLP analysis. After restrictionenzyme digestion, BsrDI (NEB, England), the samples weresubjected to electrophoresis. Four different patterns of DNAfragments were observed among 6 fungal species. Results:The DNA fragments for T. mentagrophytes var. mentagrophytes,T. mentagrophytes var. interdigitale and T. tonsuransshowed similar patterns on electrophoresis and werenot distinguishable, whereas T. rubrum, M. canis, and M. gypseum showed different patterns. Conclusion: To ourknowledge, it is the first study to introduce the analysis of thenucleotide sequence of metalloproteinase-1 enzyme tostudy differentiation in dermatophytes. Based on our results,more accurate differentiation and subtyping of T. rubrum andT. mentagrophytes var. interdigitale might be possible. Thismight contribute to better understanding of the epidemiologyand pathogenesis of dermatophyte.
참고문헌 (15)
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