Molecular Diagnosis of Pathogenic Fungi Isolated from the Skin and Mouth

Abstract

Background:  This study was conducted in the Postgraduate Laboratory at the College of Sciences at the University of Kerbala, from patients with dermatophytes ,where 115 sample were collected for the period from  December 2022 until May 2023 where 100 sample were collected ,isolated from patients with dermatophytes  which were taken from skin ,nails and hair in   Imam Al-Hassan -Almujtaba Hospital ,peace be upon him , in Karbala Governoate, and 15 samples were  taken from the mouth at the childrens teaching hospital. the  samples were directly   direct microscopic examination  and by observing the external appearance of the colonies after culturing them on medium of Saparoid dextrose agar ,the culture result showed two types of dermatophytes and atype of Candida yeast    (  Trichophyton indotinea,  Trichophyton qunickeueam and Candida albicans.) Objectives:Diagnosis of pathogenic fungi isolated from the skin and mouth molecularly because it is more accurate and specialized than the phenotypic diagnostic and evolutionary tree drawing to find the relationship between locally isolated fungi and global isolates. Materal and Methods : sampla was culture on saporoud dextrose agar,after being diagnosed phenotypic diagnosed molecular examination the DNA was extracted from the mentioned pathogenic fungi using aKorean extraction kit( Favrogene  extraction  kit for fungi ) and then the migration  was carried out  by electrophoresis,then the DNA molecule is amplified using the polymerase chain reaction(pcr).

The ITS region was adopted as the best Barcording region in the diagnosis of pathogenic fungi. The Primer pairs ITS4&ITS5 were adopted which is ageneral pair targeting the location of the gene sequence.

Result: the result of the polymerization showed unique bands  that ranged between 510-850 base pair.

Conclusion:the result of the current study showed that the genetic tree of the Dermatophytosis and Candida yeast taken from the mouth ,and after send  the results to the Gen  Bank to compare them with recorded fungi at this site and using a program MEGA(multiple alignument analysis tool6.0).the result showed a clear convergence between the local  fungi isolates compared to the rest of the  isolates present  in the tree.