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Identification of Candida species using PCR-RFLP in cancer patients in Iran
T Shokohi1, M B Hashemi Soteh, Z Saltanat Pouri
1Department of Medical Mycology and Parasitology, Sari Medical School, Mazandaran University of Medical Sciences, Sari, Iran.
Unlabelled:
Opportunistic infections caused by Non-Candida albicans. have been increasing. Traditional methods that are used to identify clinical isolates of Candida species are time-consuming and not appropriate for rapid, accurate and reliable identification.
Purpose:
To identify Candida spp isolated from cancer patients using PCR-restriction enzyme.
Materials And Methods:
Using universal primers, ITS1 and ITS4, in this study, we could amplify ITS1-5.8S-ITS2 rDNA regions at both 80 clinical isolates and 3 standard strains. The PCR products were digested with two restriction enzymes MspI and BlnI separately.
Result:
We successfully identified all isolated species using two restriction enzymes (MspI, BlnI). Candida albicans was the most common species (77.5%), followed by C. glabrata (15%), C. tropicalis (5%), C. krusei (2.5%). Although the primers and enzyme had the ability to identify C. parapsilosis, C. guilliermondii, C. dubliniensis, present isolates did not include these among identified ones.
Conclusion:
RFLP-PCR using ITSI and ITS4 primers and restriction enzyme is a rapid, easy, reliable and also applicable method in clinical laboratory for identification of medically important Candida spp.
