Identification of Candida Species Isolated from Vulvovaginal Candidiasis Patients by Polymerase Chain Reaction-Restriction Fragment Length Polymorphism (PCR-RFLP) in Yasuj Southwestern Iran

Abstract

Background: Vulvovaginal candidiasis (VVC) is a frequent infection in females at the reproductive age. Furthermore, the most common causative agent is Candida albicans yet in the recent years, the incidence of non-albicans species has increased. Objectives: The main aim of this study was the isolation and identification of various Candida species from patients with vulvovaginal candidiasis by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) in Yasuj, Iran. Methods: Three hundred and ten suspected women females vaginitis were sampled and examined. Genomic DNA was extracted from fresh colonies by phenol-chloroform glass bead methods. Polymerase chain reaction (PCR) amplification was performed based on the ribosomal DNA internal transcribed spacer (rDNA-ITS), and specific electrophoretic patterns of PCR products after digestion with MspI enzyme used for species identification. Results: The cultures were positive for 160 (51.6%) vaginal samples. Candida albicans (86.8%) was the most common species among the isolates, followed by C. glabrata (3.77%) and C. krusei (3%). Multispecies with two Candida were identified in nine patients. Conclusions: Vulvovaginal candidiasis is more prevalent among females in Yasuj and the predominant agent is C. albicans. In addition, correct identification of Candida species could play an important role in management and treatment of VVC.

Molecular identification and antifungal susceptibility profiles of Candida dubliniensis and Candida africana isolated from vulvovaginal candidiasis: A single‐centre experience in Iran

“…In this study, a preliminary molecular screening of the yeast isolates was accomplished using a polymerase chain reaction‐restriction fragment length polymorphism (PCR‐RFLP) analysis of ribosomal internal transcribed regions (ITS) 27,28 …”

“…Total genomic DNA was extracted from each isolate using the glass‐bead phenol‐chloroform method according to a previous study 29 . Purified DNA samples were then subjected to PCR‐RFLP analysis as previously described in Gharaghani et al 27 Briefly, the ITS‐rDNA regions were amplified using 0.5 μM of the fungal universal primers ITS1 and ITS4, 4 μl of DNA template, 12.5 μl of 2× Taq DNA master mix (Ampliqon) and PCR‐grade water to make a final reaction volume of 25 μl 30 . PCR cycles included initial denaturation at 94°C for 5 min, followed by 35 cycles of denaturation at 94°C for 30 s, annealing at 58°C for 30 s, extension at 72°C for 1 min and a final extension step of 7 min at 72°C.…”

“…Subsequently, the digested products were separated and visualised on a 2% (wt/vol) agarose gel stained with ethidium bromide (0.5 μg/ml) and photographed under ultraviolet light at 305 nm. Final species identification was done by comparing the obtained RFLP patterns with those of reference strains used in other studies 27,28 …”

Genetic Analysis of Candida glabrata from Candiduric Patients Using Microsatellite Length Polymorphism, Antifungal Susceptibility, and Enzymatic Profiles

“…Although Candida albicans is the most common etiological agent of candidiasis, non-albicans species have recently emerged among various systemic candidiasis forms (1). The most common non-albicans species include C. glabrata, C. tropicalis, C. krusei, and C. parapsilosis isolated from systemic and vulvovaginal candidiasis (2)(3)(4). Candiduria is one of the most common challenges in hospitalized patients, including both adults and children.…”

Prevalence of Vulvovaginal Candidiasis in Ahvaz, Southwest Iran: A Semi-Large Scale Study

“…Researchers believe that 5 -8% of adult women have RVVC (5,6,11). Several studies have shown that the frequency of VVC varies in different provinces of Iran, from 4.8 to 73% (12)(13)(14)(15)(16)(17)(18).…”