Abstract
Candidal Onychomycosis (COM) is a common nail disease which plays as sources pathogenic reservoir giving a rise to repeated candidiasis infections. This study aimed to evaluate PCR assays and phenotypic tests for identification of yeasts isolated from COM patients. The study included 100 clinically suspected patients of COM attending the main hospital and clinics in Al-Dewania province in the middle of Iraq during September 2011 to April 2012. One hundred yeast isolates were identified morphologically by CHROMagar medium. DNA was extracted from 14 representative’s isolates for accurate identification by PCR and fingerprinted by RAPD-PCR. Phenotypic examination of 100 yeasts isolates on CHROMagar revealed that these isolates were classified into 7 different species belonged to Candida form genus, PCR assay revealed that primer pair ITS1 and ITS4 was successfully amplified ITS1-5.8S-ITS2 rDNA region for 14 isolates of Candida spp. yielding a unique PCR products approximately 510-650bp in length. The results of RAPD-PCR assay showed that both primers (TAGGATCAGA and AGGTCACTGA) were genotyped 14 isolates of Candida into seven main genotypes; three of these genotypes had highly percentage of homologous (80-100%) among related isolates were studied in each Candida isolates, while the others four genotypes had 10-50% homologous. This study concluded that for accurate and prices identification must used PCR and fingerprinted by RAPD-PCR assays, the results of CHROMagar were correlated with gene expression for each Candida isolates, while the results of RAPD PCR assay were correlated with degrees similarity and difference of genotypes for Candida isolates under interest.
Highlights
Nail damage or injury, hot and humid climates (Heikkila and Stubb, 1995; Rigopoulos et al, 1998)
Phenotypic examination of 100 yeasts isolates on CHROMagar revealed that these isolates were classified into 7 different species belonged to Candida form genus, PCR assay revealed that primer pair ITS1 and ITS4 was successfully amplified ITS1-5.8S-ITS2 rDNA region for 14 isolates of Candida spp. yielding a unique PCR products approximately 510-650bp in length
Isolation and Identification of Candida spp Phenotypic examination of 100 yeasts isolates on CHROMagar revealed that these isolates were classified into 7 species belonged to Candida form genus
Summary
Nail damage or injury, hot and humid climates (Heikkila and Stubb, 1995; Rigopoulos et al, 1998). COM representative bout prolonged exposure to water, mostly infected different age and sex groups of human those followed bad hygiene, 30%-50% of all nail diseases which caused by fungi share communal showers, farmers and hard workers (Midgley et al, 1994; Garg et al, 2004). Human body included: Vagina, mouth, anal, skin and Previous studies refer that patients between 21 and 40 gastrointestinal tract. These fungi can invade skin and nail in immunocopennt and immunocompromised patients, years old were had potential Onychomycosis Identified pathogenic yeasts based on biochemical tests and phenotypic characters Such as colonial colors on CHROMagar, show reliable methods for identification but not clear cut identification most of conventional approaches were time consuming (48 to 72 h) to perform identification task delayed performed, subjected to limitations, challenges because some of biochemical tests sometime fail to identify causing agents, many reports show that 12% of tested isolates need additional treatment and 0.8% are not well identified (Beighton et al, 1995; Tarini et al, 2010)
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