Identification andDifferentiation of Trichophyton Mentagrophytes and T.rubrum by Polymerase Chain Reaction and Enzymatic Digestion
Introduction & Objective: Trichophyton rubrum and T. mentagrophytes are most common causative agents of dermatophytosis in the world. Differentiation of these species is important from the epidemiological and pathological point of view. Conventional methods including macroscopic and microscopic...
| Main Authors: | , , , , , |
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| Format: | Article |
| Language: | fas |
| Published: |
Yasuj University Of Medical Sciences
2008-07-01
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| Series: | Armaghane Danesh Bimonthly Journal |
| Subjects: | |
| Online Access: | http://armaghanj.yums.ac.ir/browse.php?a_code=A-10-39-25&slc_lang=en&sid=1 |
| Summary: | Introduction & Objective: Trichophyton rubrum and T.
mentagrophytes are most common causative agents of
dermatophytosis in the world. Differentiation of these species is
important from the epidemiological and pathological point of view.
Conventional methods including macroscopic and microscopic
morphology and biochemical tests are time-consuming (in some
cases it takes 3-4 weeks), laborious and still sometimes insufficient
to identify these agents. The aim of this study was to use
polymerase chain reaction followed by enzymatic digestion for
differentiation of these 2 species.
Materials & Methods: In this descriptive–experimental study one
hundred strains were isolated from patients with dermatophytosis.
Preliminary identification was done by morphological methods.
DNA was isolated and purified by glass-bead methods and ITS1-
5.8SrDNA-ITS2 region was amplified by PCR and the amplicon
was digested by the restriction enzyme MvaI. The products were
visualized after agarose gel electrophoresis and staining.
Differentiation of the species was based on sequence analysis and
the electrophoretic patterns.
Results: Morphological tests were not able to definitely
differentiate the two tested species, especially for isolates with
intermediate features. Using molecular methods, it was found that
45 isolates are T. rubrum and 54 are T. mentagrophytes. One
isolate was Fusarium spp. Physiological tests were confirmed the
results except for 4 isolates. It was also found that hair perforation
test is more reliable than urease test for differentiation of these two
species.
Conclusion: We found that DNA-based method, although
expensive, is a fast and reliable method for differentiation of T.
rubrum and T. mentagrophytes. The frequency of mentioned
species was almost similar in the tested isolates. The method is
recommended for differentiation of other dermatophytes. |
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| ISSN: | 1728-6506 1728-6514 |