A total of 855 yeast strains isolated from different clinical specimens, mainly nail (42%) and vulva-vagina (25%) were identified by a set of polymerase chain reaction-restriction fragment length polymorphisms (PCR-RFLP). Genomic DNA was extracted from fresh colonies using Whatman FTA Card technology. PCR assays were performed on the complete ribosomal DNA internal transcribed spacer (rDNA-ITS) region for all isolates and species identification was carried out through their specific electrophoretic profiles after digestion with the enzyme MspI. Those isolates suspected as Candida parapsilosis group were then subjected to amplification of the secondary alcohol dehydrogenase (SADH) gene and restriction digestion with NlaIII enzyme. In total, 71.1% of the strains were obtained from females and 28.9% from males. The age group of 31-40 years consisted of the highest frequency of patients with candidiasis. Candida albicans was the predominant species (58.6%) followed by C. parapsilosis (11.0%), C. glabrata (8.3%), C. tropicalis (7.0%), C. kefyr (5.8%), C. krusei (4.4%), C. orthopsilosis (2.1%), and C. guilliermondii (0.6%). A few strains of C. lusitaniae, C. rugosa, C. intermedia, C. inconspicua, C. neoformans and S. cerevisiae were isolated. We could not identify 8 (0.9%) isolates. Candida albicans remains the most frequently species isolated from Iranian patients; however, the number of non-C. albicans Candida species looks to be increasing. The simple and reliable PCR-RFLP system used in the study has the potential to identify most clinically isolated yeasts.
Background and Purpose:Candidemia is one of the most important fungal infections caused by Candida species. Infections and mortality caused by Candida species have been on a growing trend during the past two decades. The resistance of yeasts to antifungal drugs and their epidemiological issues have highlighted the importance of accurately distinguishing the yeasts at the species level. The technique applied for yeast identification should be fast enough to facilitate the imminent initiation of the appropriate therapy. Candidemia has not been studied comprehensively in Iran yet. Regarding this, the current study aimed to assess the epidemiology of candidemia at Tehran hospitals and compare the results with the previous findings.Materials and Methods:This study was conducted on 204 positive blood cultures obtained from 125 patients hospitalized in several hospitals located in Tehran, Iran, within a period of 13 months. The yeast isolation and species identification were accomplished using several phenotypic methods (i.e., production of germ tube in human serum, culture on CHROMagar Candida, and Corn meal agar containing Tween 80) and molecular methods, such as polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). In addition, unknown cases were subjected to PCR sequencing. These methods were then compared in terms of accuracy, sensitivity, and speed of identification.Results:According to the results, C. albicans (62.4%) was the most common isolate, followed by C. parapsilosis (n=36, 17.5%), C. glabrata (n=18, 8.8%), C. tropicalis (n=13, 6.3%), Trichosporon asahii (n=3, 1.5%), C. kefyr (n=2, 1.0%), C. lusitaniae (n=2, 1.0%), C. intermedia (n=1, 0.5%), C. guilliermondii (n=1, 0.5%), and C. krusei (n=1, 0.5%), respectively. Conclusion:As the findings indicated, the most common species causing candidemia were C. albicans, C. parapsilosis, and C. glabrata, respectively. Children less than one year old and people with cancer were at higher risk for candidemia, compared to other groups. Moreover, phenotypic and molecular methods resulted in the identification of 65.2% and 96.6% of the isolates, respectively. Consequently, PCR-RFLP could be concluded as a more favorable technique for species identification.
Background:In the past two decades, the incidence of fungal infections has significantly increased worldwide. Despite treatment with a broad range of antifungal agents, nosocomial candidemia is associated with high mortality rates and resistance to antifungal agents is becoming increasingly prevalent in Candida species. Therefore, detection and identification of pathogens at the species level and antifungal susceptibility testing are essential to select the appropriate antifungal therapy. The incidence of candidemia and the antifungal susceptibility patterns of the associated strains have not been extensively studied in Iran. Objectives: The current study aimed to assess the microbial epidemiology of candidemia and the antifungal susceptibility profiles of Candida isolates. Patients and Methods: Out of 5141 blood culture specimens analyzed in Iran, 48 specimens from 32 patients were yeast-positive. The isolates were precisely identified at the species level using the well-established phenotypic polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method. The yeasts were also tested for antifungal susceptibility to fluconazole, itraconazole, ketoconazole, voriconazole, and caspofungin using the broth microdilution method.
Candida glabrata is often the second most common causative agent for candidiasis following Candida albicans. Despite the importance of C. glabrata infections, few epidemiological studies have been conducted on this issue. The goal of this study was genotyping of clinical isolates of C. glabrata by multilocus sequence typing (MLST) technique for determination of the endemic prevalent genotypes and any association between isolation source and drug resistance. A total of 50 C. glabrata clinical isolates from Iran were analyzed by MLST and tested for in-vitro susceptibilities to amphotericin-B, caspofungin, fluconazole, and voriconazole according to the Clinical Laboratory Standards Institute (CLSI) M27-A4 document guidelines. Among these isolates, 16 distinct STs were identified, indicating a discriminatory power index of 0.9029. The three major sequence types (STs) were ST-59, ST-74, and ST-7 with 10, 8, and 7 isolates, respectively. Furthermore, a total of 11 new sequences were found, to which no allele numbers were assigned in the MLST database. All the isolates were susceptible to amphotericin B and caspofungin. Fluconazole resistance was shown in four isolates. Also, a sole isolate was voriconazole resistant. This study shows that the population structure of C. glabrata in Iran consists of groups closely related to the global database as well as to some new clonal clusters and STs. Regarding the high prevalence of 11 new sequences found in this study, it can be concluded that, these new alleles are among the endemic genotypes of Iran. The genotypes or STs were independent of drug susceptibility and anatomic sources.
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