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


OBJECTIVE
To identify Candida spp isolated from cancer patients using PCR-restriction enzyme.


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.


RESULTS
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.


CONCLUSIONS
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.
Shokohi
T
T
Department of Medical Mycology and Parasitology, Sari Medical School, Mazandaran University of Medical Sciences, Sari, Iran.
Hashemi Soteh
M B
MB
Saltanat Pouri
Z
Z
Hedayati
M T
MT
Mayahi
S
S
D002175:Candida / Q000145:classification* / Q000235:genetics / Q000302:isolation & purification*; D002177:Candidiasis / Q000382:microbiology*; D017931:DNA Primers / Q000235:genetics; D004271:DNA, Fungal / Q000235:genetics; D004275:DNA, Ribosomal / Q000235:genetics; D021903:DNA, Ribosomal Spacer / Q000235:genetics; D019003:Deoxyribonuclease HpaII; D015252:Deoxyribonucleases, Type II Site-Specific; D006801:Humans; D007492:Iran; D009369:Neoplasms / Q000150:complications*; D016133:Polymerase Chain Reaction / Q000379:methods*; D012150:Polymorphism, Restriction Fragment Length
D017931:DNA Primers; D004271:DNA, Fungal; D004275:DNA, Ribosomal; D021903:DNA, Ribosomal Spacer; D019003:Deoxyribonuclease HpaII; C064053:CCTAGG-specific type II deoxyribonucleases; D015252:Deoxyribonucleases, Type II Site-Specific