Identification of Candida species using PCR-RFLP in cancer patients in Iran

• Published in: Indian journal of medical microbiology Vol. 28; no. 2; pp. 147 - 151
• Main Authors: Shokohi, T, Hashemi Soteh, M. B, Pouri, Z. Saltanat, Hedayati, M. T, Mayahi, S
• Format: Journal Article
• Language: English
• Published: India Medknow Publications on behalf of Indian Association of Medical Microbiology 01.04.2010; Elsevier B.V; Elsevier Limited
• Subjects: BlnI; Candida; Candida - classification; Candida - genetics; Candida - isolation & purification; Candida albicans; Candidiasis - microbiology; Deoxyribonuclease HpaII; Deoxyribonucleases, Type II Site-Specific; DNA Primers - genetics; DNA, Fungal - genetics; DNA, Ribosomal - genetics; DNA, Ribosomal Spacer - genetics; Humans; identification; Iran; ITS region; MspI; Neoplasms - complications; Polymerase Chain Reaction - methods; Polymorphism, Restriction Fragment Length; RFLP-PCR; Iran; BlnI; ITS region; identification; MspI; RFLP-PCR; Candida
• ISSN: 0255-0857; 1998-3646
• DOI: 10.4103/0255-0857.62493

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.