Molecular Detection and Identification of Candida Species Isolates from Oral by RFLP-PCR

Abstract

In this study, 10 local isolates from a total of 50 samples of Candida sp. were collected from oral swabs of patients with oral infections in Mosul hospitals. The isolates were diagnosed based on culturing, microscopic and biochemical characteristics, and then molecular methods. The first diagnosis by culturing, microscopic and biochemical tests found the isolates were identified as Candida sp. The ITS region was amplified using universal primers (ITS4-ITS5), The PCR product was size (510-721) bp. Performing RFLP-PCR using MspI, HhaI,and EcoRI, restriction enzyme to detect and identify Candida species, the results showed the presence of the cutting sequence of MspI and HhaI enzymes in the genomic DNA content of local isolate and the absence of the sequences for the EcoRI restriction enzyme. Two Candida species were identified (C. krusei and C. the basis of size and fragment sequences then compared with sequences of standard strains from the gene bank in previous studies. Therefore, it can be observed that there is a genetic variation between the local isolates and that there are different genotypes of rDNA 5.8S have been diagnosed in 10 isolates after the cutting process with three restriction enzymes. We conclude from this study that the RFLP-PCR technique was the best in diagnosing and identifying Candida species compared with traditional methods. and we are d the genetic variation between local isolates.