Morphological and Molecular Identification of Trichophyton mentagrophytes Isolated from Dermatophytes Patients in Garmian Area

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Hana K. Maikhan
Saman M. Mohammad-Amin
Hassan M. Tawfeeq


Trichophyton mentagrophytes, PCR, RFLP, dermatophytes, ITS region.


Conventional and molecular diagnosis considered as a complementary approach for making a final decision about causative agent of all microbial population, the present study was conducted in Kalar General Hospital, Dermatology unit and research laboratory center in Garmian University. Out of thirty clinically collected specimens only five samples displayed positive dermatophytic  characters against microscopic and macroscopic as well as biochemical tests, such appearances of isolated colonies as white and brown color for both surface and reverse respectively, as well as, the colonies showed with a cottony texture which changed to powdery-granular colonies after two weeks incubation. Microscopic examination appeared numerous single-celled, spherical shaped microconidia were seen as clustered on both sides of hyphae, furthermore, multiseptate cigar shaped macroconidia and spiral hyphae were seen during the formation of granular colonies, biochemically analysis showed positive for urease and hair perforation testes. Molecular identification carried out via conventional PCR protocol by using the primers set ITS1 and ITS4 which result in 700bp in agarose gel electrophoresis for all isolates. PCR-RFLP carried out by means of using BstN1 digestion enzyme revealed four separated pattern bands 250,180,150 and 120bp. Sequencing of the ITS region in one of the isolated species revealed  that  similarity about 86% with Trichophyton mentagrophytes ATCC11481

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AHMADI, B.; MIRHENDI, H.; SHIDFAR, M.R.; NOURIPOUR-SISAKHT, S. JALALIZAND, N.; GERAMISHOAR, M. AND SHOKOOHI, G.R. 2015. A comparative study on morphological versus molecular identification of dermatophyte isolates. J Mycol Med, 25(1):29-35.
DE HOOG, G.S.; GUARRO, J.; GENE, J. AND FIGUERAS, M.J. 2000. Atlas of Clinical Fungi 2nd edition. Centraalbureau voor Schimmelcultures, Utrecht, The Netherlands.
DHIEB, C.; ESSGHAIER, B.; EL EUCH, D. AND SADFI-ZOUAOUI, N. 2014. Phenotypical and Molecular Characterization of Microsporum canis Strains in North-Tunisia. Polish Journal of Microbiology, 63(3):307-315.
ELAVARASHI, E.; KINDO, A. J.AND KALYANI, J. (2013). Optimization of PCR–RFLP Directly from the Skin and Nails in Cases of Dermatophytosis, Targeting the ITS and the 18S Ribosomal DNA Regions. Journal of Clinical and Diagnostic Research, 7(4): 646-651.
FOROBES, B.A.; SAHM, D.F. AND WEISSFELD, A.C. 1998. Dignostic Microbiology. 10th edition., Mosby, Inc. London.
GHOJOGHI, A.; FALAHATI, M.; PAGHEHM A.S.; ABASTABAR, M.; GHASEMI, Z.; ANSARI, S.; FARAHYAR, S. AND ROUDBARY, M. 2015. Molecular Identification of Epidemiological Aspect of Dermatophytosis in Tehran, Iran. J. Research in Molecular Medicine, 3:11-16.
GRASER, Y.; SCOTT, J. AND SUMMERBELL, R. 2008. The new species concept in dermatophytes – a polyphasic approach. Mycopathologia, 166, 239–256
JACKSON-FISHER, A.J., BURMA, S., PORTNOY, M., SCHNEEWEIS, L.A., COLEMAN, R.A., MITRA, M., CHITIKILA, C. AND PUGH, B.F., 1999. Dimer dissociation and thermosensitivity kinetics of the Saccharomyces cerevisiae and human TATA binding proteins. Biochemistry, 38(35),11340-11348.
KANBE, T. 2008. Molecular approaches in the diagnosis of dermatophytosis. Mycopathologia, 166(5-6), 307.
KANBE,T.; SUZUKI, Y.; KAMIYA, A.; MOCHIZUKI, T.; KAWASAKI, M.; FUJIHIRO, M. AND KIKUCHI, A. 2003. Species-identification of dermatophytes Trichophyton, Microsporum and Epidermophyton by PCR and PCR-RFLP targeting of the DNA topoisomerase II genes. Journal of Dermatological Science,33, 41-54.
LI, H.C., BOUCHARA, J.P., HSU, M.M.L., BARTON, R., SU, S. AND CHANG, T.C. 2008. Identification of dermatophytes by sequence analysis of the rRNA gene internal transcribed spacer regions. Journal of medical microbiology, 57(5), 592-600.
MAIKHAN, H.K. AND MOHAMMAD-AMIN, S.M. 2017. Isolation, Biochemical Identification and Antioxidant Activity of Locally Isolated Lactobacillus spp in Garmian Area. Eurasian Journal of Science & Engineering, 3(1). 128-136
NENOFF P, HERRMANN J, GRASER Y. TRICHOPHYTON MENTAGROPHYTES SIVE INTERDIGITALE?. 2007. A dermatophyte in the course of time. J Dtsch Dermatol Ges, 5: 198–202.
NENOFF, P., ERHARD, M., SIMON, J.C., MUYLOWA, G.K., HERRMANN, J., RATAJ, W., GRÄSER, Y. (2013). MALDI-TOF mass spectrometry – a rapid method for the identification of dermatophyte species. Med. Mycol, 51, 17–24.
POLURI, L.V., INDUGULA, J.P. AND KONDAPANENI, S.L. (2015). Clinicomycological study of dermatophytosis in South India. Journal of laboratory physicians, 7(2), p.84.
Refai, M.; El-Yazid, H. A. and Al- Hariri, M. 2013. Monograf on Dermatophytes. A guide for isolation and identification of dermatophytes, diseases and treatment. Cairo. Egypt.
REZAEI-MATEHKOLAEI, A., MAKIMURA, K., SHIDFAR, M.R., ZAINI, F., ESHRAGHIAN, M.R., JALALIZAND, N., NOURIPOUR-SISAKHT, S., HOSSEINPOUR, L. AND MIRHENDI, H. 2012. Use of single-enzyme PCR-restriction digestion barcode targeting the internal transcribed spacers (ITS rDNA) to identify dermatophyte species. Iranian journal of public health, 41(3),82.
SINGH, S. AND BEENA, P.M. 2003. Profile of dermatophyte infections in Baroda. Indian Journal of Dermatology, Venereology, and Leprology, 69(4), 281-283.
SURENDRAN, K.A.K., BHAT, R.M., BOLOOR, R., NANDAKISHORE, B. AND SUKUMAR, D., 2014. A clinical and mycological study of dermatophytic infections. Indian journal of dermatology, 59(3),.262.
SYMOENS, F.; JOUSSON, O.; PLANARD, C.; FRATTI, M.; STAIB,P.; MIGNON,B. AND MONOD, M. 2011. Molecular analysis and mating behaviour of the Trichophyton mentagrophytes species complex. International Journal of Medical Microbiology, 301, 260–266.
YU, J.; WAN, Z.; CHEN, W.; WANG, W. AND LI, R. 2004. Molecular typing study of the Microsporum canis strains isolated from an outbreak of tinea capitis in a school. Mycopathologia 157(1): 37–41.