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2014 ; 26
(3
): 338-42
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Identification of dermatophytes by polymerase chain reaction-restriction fragment
length polymorphism analysis of metalloproteinase-1
#MMPMID24966633
Jung HJ
; Kim SY
; Jung JW
; Park HJ
; Lee YW
; Choe YB
; Ahn KJ
Ann Dermatol
2014[Jun]; 26
(3
): 338-42
PMID24966633
show ga
BACKGROUND: Transgenic research on metalloproteinase-1 is an emerging field in
the area of plant molecular biology. The new method reported here can similarly
be applied in fungal molecular biology to identify different dermatophytes. Our
method is more accurate than traditional methods such as molecular analyses.
OBJECTIVE: To identify Trichophyton rubrum, T. mentagrophytes var.
mentagrophytes, T. tonsurans, T. mentagrophytes var. interdigitale, Microsporum
canis and M. gypseum, by using the restriction fragment length polymorphism
(RFLP) analysis and polymerase chain reaction (PCR) to detect polymorphisms in
the metalloproteinase-1 gene (MEP1). METHODS: From each fungal strain, we
isolated genomic DNA and performed PCR to amplify the region coding for
metalloproteinase-1. Primers for the metalloproteinase-1 gene were designed based
on the sequence in NCBI GenBank. Subsequently, we purified the amplified PCR
product and performed RFLP analysis. After restriction enzyme digestion, BsrDI
(NEB, England), the samples were subjected to electrophoresis. Four different
patterns of DNA fragments were observed among 6 fungal species. RESULTS: The DNA
fragments for T. mentagrophytes var. mentagrophytes, T. mentagrophytes var.
interdigitale and T. tonsurans showed similar patterns on electrophoresis and
were not distinguishable, whereas T. rubrum, M. canis, and M. gypseum showed
different patterns. CONCLUSION: To our knowledge, it is the first study to
introduce the analysis of the nucleotide sequence of metalloproteinase-1 enzyme
to study differentiation in dermatophytes. Based on our results, more accurate
differentiation and subtyping of T. rubrum and T. mentagrophytes var.
interdigitale might be possible. This might contribute to better understanding of
the epidemiology and pathogenesis of dermatophyte.
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