Use of PCR-RFLP and PCR-HWP1 for identification of Candia species isolated from cystic fibrosis patients

Due to the predisposing conditions in patients with cystic fibrosis (CF) caused by defective mucociliary clearance facilitates of colonization and invasion with Candida species has dramatically increased. Traditional methods for identification problems are imminent and are time-consuming. Therefore, molecular techniques utilizing amplification of target DNA provide quick and precise methods for diagnosis and identification of Candida species. Therefor, the aim of current study was identification of the most medically common isolated Candida species from the air way of CF patients by PCR-RFLP and amplification of HWP1 gene. A total of 42 CF patients presenting symptoms referred to pediatric respiratory diseases research center were screened for the presence of Candida spp. The isolates initially were phenotypically identified and confirmed by molecular approaches based on restriction fragment length polymorphism (PCR-RFLP) for discrimination of C. albicans of non albicans and amplification of HWP1 gene for discrimination of C. albicans from C. dubliniensis and C. africana was conducted. Results show that C. albicans was the most frequently isolated species (83.8%) followed by non-albicans included C. parapsilosis (7.1%), C. glabrata (3.2%), and C. tropicalis (3.2%). The restriction patterns of each Candida species were perfectly specific. Since MspI is not able to discriminate between three morphological related species, C. albicans, C. dubliniensis and C. africana, we used PCR amplification of hwp1 gene, which (7.1%) species from C. albicans identified as C. dubliniensis, however C. africana strains were not found. The present study found that C. albicans as predominant species isolated from the CF patients. It can be concluded that molecular diagnostic methods are reliable and would be useful for identification of medically important Candida species in clinical samples. Therefore considerable attention has been paid to prevention and treatment of microbial growth, which has resulted in improvement of patient management.