Evaluation of expression of CDR1, CDR2 genes in clinical isolates of Candida albicans resistant and susceptible to fluconazole in patients under chemotherapy in Ahvaz

Introduction: Nowadays candidiasis is being into new treatment problem. Resistance to azole drugs during prolonged use and poor response were seen to treatment in candidiasis. The effect of azole drugs on Candida albicans isolates is controlled by several genes, including the genes. They have different molecular mechanisms to resistance azoles drug.In this study, 12 susceptible isolates and 12 isolates of fluconazole resistant Candida albicans were evaluated by qPCR, as a rapid and sensitive assay, to find overexpression of the mentioned genes along with drug sensitivity test.Materials and Methods: The Candida species were obtained by swabbing the oral mucosa of cancer patients in Ahvaz. The isolates were identified by morphotyping and molecular methods. Then, RNA extraction and cDNA synthesis was carried out in 24 Candida albicans isolated from patients. Antifungal susceptibility tests were conducted using micro dilution broth method. MICs were determined after 24 h of incubation at 35 °C and interpreted according to (EUCAST) breakpoints. Primer blast software was used for primers designed for CDR1, CDR2 and ACT1 (internal control) in RT- qPCR.RT- qPCR amplification of reference genes and target genes was performed by using ABI Step One®.Results: In total, 74 Candida isolates were diagnosed from patients. After 24h 21.6% of the isolate were resistant to fluconazole, 10.8% of the isolate were dose-dependent and 67.7% of the isolate were sensitivite. Resistant isolates were identified significantly with over expression in CDR1 gene. Compared CDR2 gene between clinical isolates of fluconazole-sensitive and resistant Candida albicans, did not show significant overexpression.Conclusion: It is important to identify the mechanism of resistance to azole drugs and to know the epidemiology of Candida isolates in clinical laboratories. Rapid molecular methods such as RT-qPCR to identify mechanism of resistance to fluconazole in under chemotherapy patients can significantly affect treatment management of these patients.

کلیدواژه ها:

Oral candidiasis ، Chemotherapy ، MIC ، RT- qPCR ، CDR1 ، CDR2