Cryptococcus and cryptococcosis in Iran during 1969-2019: A systematic review and meta-analysis.

OBJECTIVE: Limited data are available on the epidemiology and etiology of cryptococcal infections in the Middle East. We aimed to conduct the systematic review and meta-analysis to summarize the epidemiological data on prevalence of Cryptococcus species complexes in trees and their surroundings, bird guano and secretions, animals, and highlight the reported episodes of cryptococcosis in Iran. MATERIALS AND METHODS: Twelve databases, including PubMed, Science Direct, Scopus, Proquest, Google Scholar, Embase, and the ISI Web of Science, as well as the national databases, from January 1969 to October 2019 were searched. Furthermore, gray literature (e.g., thesis, congress abstracts) was evaluated using Iran Doc and www.thesis. RESEARCH: ac.ir. Search process was accomplished on English or Persian language articles using the following keywords: "Cryptococcus", "Cryptococcosis", "invasive fungal infection", "Humans", "Birds", "Pigeon", "Animals", "Tree", "Eucalyptus", and "Iran", both alone and in combination. RESULTS: Overall 36 studies were eligible regarding Cryptococcus and cryptococcosis in Iran. The total prevalence rates of Cryptococcus species in the tree was 4.7% (95% CI: 2.3-7.8), and in bird guano was 20.4% (95% CI: 10.7-32.2). Cryptococcosis in animal, and human were 1.7% (95% CI: 0.01-5.1), and 2.8% (95% CI: 0.7v6.1), respectively. The highest prevalence of Cryptococcus in the trees (14.6%), and bird guano (89.4%) in Khorasan, animals (8.9%) in Chaharmahal and Bakhtiari, and human (4.4%) in Mazandaran provinces were reported. CONCLUSIONS: Given the significant risk of Cryptococcus species for susceptible humans, mainly HIV-infected patients, it seems quite necessary to adopt concrete preventive strategies to pinpoint the environmental habitats of this yeast.

Molecular identification and antifungal susceptibility testing of Candida species isolated from dental plaques.

OBJECTIVE: The aim of the present study was to provide insight into the prevalence and susceptibility profiles of Candida species isolated from the dental plaque of Iranian immunocompetent patients. As a biofilm, Candida species are responsible for several disorders common to the oral cavity including gingivitis, dental caries, periodontitis, and the less common severe systemic infections specifically in immunosuppressed individuals. METHOD: PCR-RFLP was performed to identify yeasts isolated from the dental plaques of 40 immunocompetent patients. Moreover, antifungal susceptibility testing was performed in according to CLSI guidelines (M27-A3). RESULTS: Among 40 yeasts isolated from the dental plaques of immunocompetent patients, Candida albicans was the most common species (92.5%), followed by P. kudriavzevii (7.5%). It is the first isolation of P. kudriavzevii from dental plaques and the first evaluation of antifungal effect of the new imidazole, luliconazole and echinocandins against these samples worldwide. Luliconazole, voriconazole, amphotericin B and anidulafungin showed the best activity with the lowest geometric mean (GM) 0.03, 0.06, 0.08 and 0.09µg/ml, respectively, followed by miconazole (0.14µg/mL), caspofungin (0.24µg/mL) fluconazole (0.38µg/mL) and itraconazole (0.5µg/mL). CONCLUSION: The current study demonstrated luliconazole and echinocandins displayed excellent activity against all Candida isolates from dental plaques, presenting promising and potent alternative for all oral Candidiasis.

Voriconazole associated mucormycosis in a patient with relapsed acute lymphoblastic leukemia and hematopoietic stem cell transplant failure: A case report.

The patients with hematologic malignancies and hematopoietic stem cell transplantation (HSCT) recipients are at high risk for invasive fungal diseases (IFDs) mainly due to the severe and prolonged neutropenia related to high-dose chemotherapy. Voriconazole prophylaxis is recommended for possible IFDs. Mucormycosis is a fulminant infection, which may occur after voriconazole prophylaxis for invasive aspergillosis in immunocompromised hosts. Here, we report mucormycosis after 4 months of voriconazole prophylaxis in a young patient with relapsed acute lymphoblastic leukemia and hematopoietic stem cell transplant failure and discuss the clinical manifestation, imaging, laboratory findings and therapeutic regimens. Clinician's awareness of this entity and timely diagnosis using conventional and molecular methods are the promising approach for the management of this devastating infection.

Fulminant mucormycosis of maxillary sinuses after dental extraction inpatients with uncontrolled diabetic: Two case reports.

Mucormycosis is a rare but fulminant opportunistic fungal infection, which occurs most often in diabetic and immunocompromised patients. Dental extractions may create a portal of entry for the fungal infection. The mucormycosis may be the original cause of the pain and can be misdiagnosed as dental pain. In this paper, two cases of mucormycosis are reported after dental extractions and successfully treated with amphotericin B (case #1) and combined with posaconazole (case #2). The two cases we describe exemplify the fulminant mucormycosis of maxillary sinuses after dental extraction inpatients with uncontrolled diabetic support the findings that this predisposing condition created a suitable environment for the Mucorales growth. These case reports emphasize early recognition and urgent treatment of mucormycosis is necessary to prevent the spread of infection Therefore, dental surgeons and healthcare practitioners should become familiar with mucormycosis.

In vitro antitumor activity of patulin on cervical and colorectal cancer cell lines.

BACKGROUND AND PURPOSE: Patulin is a mycotoxin produced by some molds, especially Aspergillus and Penicilium, and is responsible for mycotoxicosis in animals and humans.There is still not very detailed data about the anti-cancer potency of patulin, but some reports demonstrated that it induces cellular apoptosis and toxicity. MATERIALS AND METHODS: To determine the efficacy of patulin as a therapeutic strategy for cervical and colorectal cancers, we investigated its effects on HeLa,SW-48, and MRC-5 cell lines. Cell lines were exposed to various concentrations of patulin (i.e., 0.5, 1, 2, and 4 µM), then using methyl thiazolyl tetrazolium (MTT) and bromo-2'-deoxyuridine (BrdU) assays, the rates of apoptosis and cell viability were determined. RESULTS: The obtained results showed a significant reduction in cell viability and apoptosis induction in a dose-dependent manner. Among all the cell lines, the highest growth inhibition rate was obtained at the 4 µM concentration of patulin. CONCLUSION: Our results suggested that patulin could significantly decrease tumor growth in human cervical and colorectal cancer models.

Characterization of clinically important dermatophytes in North of Iran using PCR-RFLP on ITS region.

OBJECTIVE: Dermatophytosis is considered as one of the major public health problems in the world. Accurate identification of the dermatophytes species is important for recognition of the environment and animal sources of infection. In this survey, 1500 clinical specimens taken from outpatients suspected of dermatophytosis and they were examined by direct microscopy and culture. MATERIALS AND METHODS: In total, three hundred and twelve dermatophytes were isolates recovered in culture, they were identified using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method based on the ribosomal DNA internal transcribed spacer (ITS) regions. RESULTS: Tinea pedis was the most common clinical form (32.1%) followed by tinea cruris (24.4%), tinea manuum (16.4%), tinea unguium (8.3%), tinea corporis (7.3%), tinea faciei (6.4%), and tinea capitis (5.1%). Trichophyton interdigitale was the most frequent isolate (38.2%), followed by Trichophyton rubrum (29.8%), Epidermophyton floccosum (16.6%), Trichophyton tonsurans (14.8%) and Microsporum canis (0.6%). The frequency of dermatophytosis was higher in males than in females and in the age-group of 21-30 years. CONCLUSION: Our finding indicated that the incidence of dermatophytosis caused by anthropophilic dermatophytes in Mazandaran province is increasing. Also, this study provides valuable data for the prevention and control of dermatophytosis in the southern coast of the Caspian Sea.

Development of a high-resolution melting analysis assay for rapid and high-throughput identification of clinically important dermatophyte species.

Accurate identification of dermatophyte species is important both for epidemiological studies and for implementing antifungal treatment strategies. Although nucleic acid amplification-based assays have several advantages over conventional mycological methods, a major disadvantage is their high cost. The aim of this study was to develop a rapid and accurate real-time PCR-based high-resolution melting (HRM) assay for differentiation of the most common dermatophyte species. The oligonucleotide primers were designed to amplify highly conserved regions of the dermatophyte ribosomal DNA. Analysis of a panel containing potentially interfering fungi demonstrated no cross reactivity with the assay. To evaluate the performance characteristics of the method, a total of 250 clinical isolates were tested in comparison with the long-established PCR-RFLP method and the results were reassessed using DNA sequencing, as the reference standard method. The assay is able to type dermatophytes using normalised melting peak, difference plot analysis or electrophoresis on agarose gel methods. The results showed that, in comparison to PCR-RFLP, the developed HRM assay was able to differentiate at least 10 common dermatophytes species with a higher speed, throughput and accuracy. These results indicate that the HRM assay will be a useful sensitive, high throughput and cost-effective method for differentiating the most common dermatophyte species.

Hyphal wall protein 1 gene: A potential marker for the identification of different Candida species and phylogenetic analysis.

BACKGROUND AND PURPOSE: Hyphal wall protein 1 (HWP1) is an important adhesin which usually is expressed on the germ tube and hyphal surface produced by different Candida species. The hyphal wall protein-coding gene (HWP1) was evaluated as a novel identification and phylogenetic marker in Candida tropicalis, C. orthopsilosis, C. parapsilosis and C. glabrata. MATERIALS AND METHODS: Initially, four specific primer pairs were designed, and the target was amplified and finally sequenced. A total of 77 Candida isolates from four different species were included in the study. Consensus sequences were used for the evaluation of phylogenetic tree using the CLC Genome Workbench, GENEIOUS, and MEGA softwares and the levels of nucleotide and amino acid polymorphism were assessed. RESULTS: According to the results, the specific amplified fragments of HWP1 gene were useful for the differentiation of four species. Intra-species variation was observed only in C. tropicalis with two DNA types. The phylogenetic tree of Candida species based on the HWP1 gene showed consistency in topology with those inferred from other gene sequences. CONCLUSION: We found that HWP1 gene was an excellent marker for the identification of non-albicansCandida species as well as the phylogenetic analysis of the most clinically significant Candida species.

Clinical features, diagnosis, and outcomes of rhino-orbito-cerebral mucormycosis- A retrospective analysis.

BACKGROUND AND PURPOSE: Rhino-orbito-cerebral mucormycosis (ROCM) is a rare disease with acute and fulminant manifestation. This infection is associated with high morbidity and mortality rates. Herein, we reviewed the manifestations, underlying conditions, medical treatments, and surgical interventions in ROCM patients admitted to a tertiary referral center in northern Iran over a seven-year period. MATERIALS AND METHODS: In a retrospective analysis, 15 cases of ROCM were identified from 2007 to 2013 in Bu Ali Sina Hospital, Sari, Iran. All the ROCM cases were clinically diagnosed and confirmed by histopathological and/or mycological examination. The relevant demographic data, clinical, ophthalmic, and neurologic manifestations, underlying conditions, medical treatments, and surgical interventions were recorded and analyzed. RESULTS: The mean age of the patients was 54±11 years (age range: 28-70 years); 26.7% of the patients were male and 73.3% female (male: female ratio of 1: 2.7). Uncontrolled diabetes was noted in at least 86.7% (13/15) of the cases. The maxillary sinuses were the most frequently involved sites (66.7% of the cases) followed by the ethmoid sinus. Amphotericin B in combination with surgical debridement was used in the treatment of 80% of the cases. Furthermore, 73.3% of the patients who were diagnosed early and underwent medical and extensive surgical debridement of the infected tissues survived. CONCLUSION: Uncontrolled diabetes mellitus is considered to be the main predisposing factor for ROCM. To prevent and reduce mortality rate of this acute disease, early diagnosis based on clinical findings and biopsy is recommended.

In vitro activity of five antifungal agents against Candida albicans isolates, Sari, Iran.

BACKGROUND AND PURPOSE: Candidaalbicans is the most common causative agent of candidiasis. Candidiasis management is dependent on the immune status of the host, severity of disease, and the choice of antifungal drug. Antifungals, specifically triazoles, are widely administered for the treatment of invasive fungal infections. Herein, we aimed to evaluate the invitro susceptibility of C.albicans isolates to fluconazole (FLZ), itraconazole (ITZ), voriconazole (VRZ), amphotericin B (AMB), and Caspofungin (CAS). MATERIALS AND METHODS: A total of 44 clinical strains of C.albicans were collected from 36 patients admitted to four hospitals in Mazandaran Province, Iran. The invitro antifungal susceptibility testing was performed based on the Clinical and Laboratory Standards Institute methods. RESULTS: Generally, 34 isolates were susceptible to all the five antifungal drugs, while four isolates were susceptible or susceptible dose-dependent (SDD) and six isolates were SDD or resistant to these antifungals. The lowest minimum inhibitory concentration (MIC; 0.016 µg/ml) belonged to AMB and the highest MIC was for FLZ )16 µg/ml). The lowest MIC (50 0.063 µg/ml) was related to ITZ and the lowest MIC (90 0.25 µg/ml) pertained to CAS, in addition , the highest MIC (50 1 µg/ml) and MIC (90 4 µg/ml) were for FLZ. Four of the isolates showed resistance to both FLZ and VRZ, separately, and five isolates were resistant to ITZ. Caspofungin showed potent activity against more than %95 of the C.albicans isolates. CONCLUSION: Overall, we reported %9.1 resistance to FLZ and VRZ ,%11.3 resistance to ITZ and AMB, and %4.6 resistance to caspofungin .Our finding is in agreement with previous observations proposing that C.albicans isolates develop resistance to some antifungal drugs such as FLZ since they are widely used as prophylaxis.

A real time PCR assay on blood for diagnosis of invasive candidiasis in immunocompromised patient.

BACKGROUND AND PURPOSE: Invasive candidiasis (IC) is a significant cause of morbidity and mortality in patients with hematologic disorders and bone marrow transplant recipients. Rapid, specific and sensitive test for the timely accuracy in immunocompromised patients to reduce mortality rates and prevent IC progress is necessary. We established a real-time PCR assay on blood for the diagnosis and differentiation of the causative Candida species. MATERIALS AND METHODS: Whole blood samples were collected twice, from 72 patients for Real Time PCR and blood culture assays. The primers and hybridization probes were designed to potentiate the specific sequence of 18S rRNA genes using Light Cycler system and Fluorescence Resonance Energy Transfer (FERT). The patients with hematologic malignancies and bone marrow transplant recipients were evaluated for IC based on the revised European Organization for Research and Treatment of Cancer/ Mycoses Study Group (EORTC/MSG) criteria. RESULTS: From 2009 to 2011, 72 patients with hematologic malignancies and bone marrow transplant recipients were evaluated for IC. The female to male ratio was 27:45; the mean age was 32.1 years. The most common malignancy in this patient was acute myeloid leukemia (AML) (27.8%) and acute lymphoblastic leukemia (ALL) (26.4%). Out of 72 patients, 11 patients (15.3%) had positive real time PCR /probe results. Based on the melting temperature (Tm) analysis, 5 (45.4%) C. krusei, 3 (27.2%) C. tropicalis, 2 (18.1%) C. parapsilosis and 1 C. albicans (9%) were identified. According to the revised EORTC / MSG, 1 patient (9%) and 10 patients (91%) were defined as proven and possible groups of IC, respectively. The mortality rate in proven and possible IC patient was found 54.5%. CONCLUSION: The established Real-time PCR/FRET probe assay is an appropriate diagnostic tool for the detection of Candida species DNA and the management of patients suffering from hematologic malignancies and bone marrow recipient are at risk for IC.

Detection of fungi by conventional methods and semi-nested PCR in patients with presumed fungal keratitis.

BACKGROUND AND PURPOSE: Fungal keratitis is a suppurative, ulcerative, and sight-threatening infection of the cornea that sometimes leads to blindness. The aims of this study were: recuperating facilities for laboratory diagnosis, determining the causative microorganisms, and comparing conventional laboratory diagnostic tools and semi-nested PCR. MATERIALS AND METHODS: Sampling was conducted in patients with suspected fungal keratitis. Two corneal scrapings specimens, one for direct smear and culture and the other for semi- nested PCR were obtained. RESULTS: Of the 40 expected cases of mycotic keratitis, calcofluor white staining showed positivity in 25%, culture in 17.5%, KOH in 10%, and semi-nested PCR in 27.5%. The sensitivities of semi-nested PCR, KOH, and CFW were 57.1%, 28.5%, and 42% while the specificities were 78.7%, 94%, and 78.7%, respectively. The time taken for PCR assay was 4 to 8 hours, whereas positive fungal cultures took at least 5 to 7 days. CONCLUSION: Due to the increasing incidence of fungal infections in people with weakened immune systems, uninformed using of topical corticosteroids and improper use of contact lens, fast diagnosis and accurate treatment of keratomycosis seems to be essential. Therefore, according to the current study, molecular methods can detect mycotic keratitis early and correctly leading to appropriate treatment.

In vitro activity of econazole in comparison with three common antifungal agents against clinical Candida strains isolated from superficial infections.

BACKGROUND AND PURPOSE: Candida species are the most common organisms involved in superficial fungal infections, worldwide. Although econazole is among the most frequently used topical formulations for the treatment of candidiasis, no information is available regarding the susceptibility profiles of Candida species in Iran. MATERIALS AND METHODS: In vitro susceptibility of 100 clinical Candida isolates belonging to 6 species from superficial candidiasis of Iran towards to econazole was compared with three other common antifungal agents including itraconazole, fluconazole, and miconazole. Minimum inhibitory concentrations (MICs) values were analyzed according to the Clinical and Laboratory Standards Institute (CLSI) M38-A3 document. All isolates were previously identified to the species level, using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) on ITS region. RESULTS: The MIC of econazole, itraconazole, miconazole, and fluconazole were within the range of 0.016-16, 0.032-16, 0.016-16, and 0.25-64 µg/ml, respectively. In general, econazole and miconazole were more active against Candida isolates, compared to the other two agents. CONCLUSION: The present study demonstrated that for Candida albicans isolates, miconazole and econazole had the best effect, but in non-albicans Candida species, itraconazole and miconazole displayed more activity than other antifungal agents.

Study on fungi in archives of offices, with a particular focus on Stachybotrys chartarum.

OBJECTIVE: This study aimed at evaluating fungi in archives of different offices in Sari city the capital of Mazandaran, a northern province of Iran, with a particular focus on Stachybotrys chartarum. MATERIALS AND METHODS: The samples were collected from twenty archives of offices and controls (n=7) using a SKC single-stage impactor which draws air at 20L/min (100L) and impacts the sampled material onto Petri dishes containing malt extract agar (MEA) (n=22) and also cellulose agar (CA) (n=22). Surface samples were also collected by pressing a sterile cotton swab on different areas of archives and cultured on MEA and CA. The grown fungi were identified by standard mycological techniques. The counted fungal colonies were converted to CFUs per cubic meter. RESULTS: In indoor air of archives, Cladosporium spp (25.1%), Aspergillus spp (22.9%) and Penicillium spp (22.9%) had the most frequencies. Stachybotrys chartarum (7.9%) was the fourth most common fungus isolated from the surface samples. Cladosporium spp had the highest total CFU concentration in indoor air of archive samples (1227/m(3)). Stachybotrys chartarum was recovered from surface collected samples of 4 archives of offices on CA. Out of the 22 rooms of archives, 45.4%, 45.4% and 9.1% had concentration level < 170 CFU/m(3), > 170 < 560 CFU/m(3) and > 560 < 1000 CFU/m(3), respectively. CONCLUSION: The results of our study have shown the high concentration levels of airborne fungi in some archives of offices that might put the workers at risk from respiratory diseases.

Epidemiology and molecular characterization of Cryptococcus neoformans isolated from pigeon excreta in Mazandaran province, northern Iran.

OBJECTIVE: The aims of this study were to verify the presence of Cryptococcus neoformans in pigeon excreta in Mazandaran province, Iran, to identify the varieties of the C. neoformans isolates using D1/D2 and IGS sequencing, and determining the presence of the two mating types: α and a. MATERIALS AND METHODS: Four hundred pigeon droppings samples were collected from 15 different cities in Mazandaran province over a period of 1 year (February 2010-March 2011). Identification of C. neoformans was determined based on growing brown colonies on Niger seed agar (NSA) and biochemical characteristics. We used MATα and MATa specific primers for determining mating type and sequence analysis of the D1/D2 and intergenic spacer regions were done. RESULTS: Out of 400 samples, 20 samples (5%) were positive for C. neoformans and all of these isolates were α mating types. Sequence analysis of polymerase chain reaction (PCR) amplicons of D1/D2 regions revealed that all of the isolates were C. neoformans var. grubii except two isolates that were C. neoformans var. neoformans. CONCLUSION: Our results reinforced that the pigeon excreta is a favorable environment rich in nitrogen and supports the growth of C. neoformans and the pigeon could play an important role in spread of this organism.

Study on fungal flora of tap water as a potential reservoir of fungi in hospitals in Sari city, Iran.

OBJECTIVE: The aim of the present investigation was to evaluate the fungal flora of tap water from university hospitals of Sari city, Iran. MATERIALS AND METHODS: During a 1-year period, 240 water samples were collected from four university hospitals. All water samples were collected in sterile polystyrene bottles. A volume of 100ml of the samples passed through sterile 0.45-micrometer filters. The filters were placed directly on malt extract agar and incubated at 27°C for 3 to 7 days. Routine mycological techniques were applied to identification of grown fungi. RESULTS: Out of 240 plates, 77.5% were positive for fungal growth. Twelve different genera were identified. Aspergillus (29.7%), Cladosporium (26.7%) and Penicillium (23.9%) were the most common isolated. Among Aspergillus species, A. flavus had the highest frequency. Highest colony counts were found in autumn. Aspergillus predominated in autumn, Cladosporium in winter and spring and Penicillium in summer. CONCLUSION: The results of our study showed that hospital water should be considered as a potential reservoir of fungi particularly Aspergillus.

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

UNLABELLED: 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. PURPOSE: To identify Candida spp isolated from cancer patients using PCR-restriction enzyme. MATERIALS AND 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. RESULT: 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. CONCLUSION: 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.

Identification of Malassezia species isolated from Iranian seborrhoeic dermatitis patients.

BACKGROUND AND OBJECTIVES: In recent years, the genus Malassezia has come to be considered important in the etiology of seborrhoeic dermatitis (SED). The aim of present study was identification of Malassezia species on the lesions of Iranian SED patients. METHODS: 100 patients with SED were enrolled in the study. The patients were evaluated both clinically for the severity of SED and microscopically for the presence of the yeast Malassezia. Diagnosis of Malassezia was made after the yeast Malassezia was microscopically observed on skin scales stained with methylene blue. All samples were also cultivated on Leeming and Notman and Sabouraud's dextrose agar culture media. The agar plates were incubated at 32 degrees C for 2 weeks and evaluated for the existence of growth every day for one week. Identification of isolated yeast was based on morphological and physiological characteristics. RESULTS: From 100 patients with SED, 60% were female. The age range was 12-65 years with median 27.3 years. The highest prevalence of SED was seen in 20-29 years age group. 59% and 41% of patients had local and generalized lesions, respectively. 58% of patients showed lesion on scalp. Microscopic examination of skin scales was positive in 100% of SED lesions. 96% of patients showed more than 1-3 yeasts in each microscopic field whereas only 4% patients showed 1-3 yeasts in whole slide. Totally, 77% of the specimens yielded Malassezia in culture. Malassezia globosa was the most commonly isolated Malassezia species (55.8%). Malassezia globosa had also most frequencies on scalp and face lesions. Malassezia furfur had most frequency on trunk lesions. CONCLUSION: The results of our study showed high recovery rate of Malassezia species on lesions of patients with SED. So it might be playing a causative role in the etiology of this disease.

Distribution of Malassezia species in patients with pityriasis versicolor in Northern Iran.

PURPOSE: Malassezia yeasts are globally distributed agents of pityriasis versicolor and are implicated in the pathogenesis of seborrhoeic and atopic dermatitis. The aim of this study is to identify the Malassezia species obtained from pityriasis versicolor patients, using morphological, biochemical, physiological as well as Polymerase Chain Reaction-Restriction Fragment Length Polymorphism (PCR-RFLP) methods. MATERIALS AND METHODS: The identification of Malassezia species is performed according to microscopic features and physiological characteristics, including catalase reaction and Tween assimilation tests. The DNA is extracted from cultured Malassezia using the glass bead, phenol-chloroform method. The internal transcribed spacer 1(ITS1) region is amplified and there is restricted digestion of the PCR products with two enzymes Cfo I and Bst F5I. RESULTS: The most commonly isolated species is M. globosa (47.6%). RFLP analysis of the PCR products of the ITS1 region is in complete agreement with those from the DNA sequences of the internal transcribed spacer (ITS) 1 region and the biochemical tests. CONCLUSION: Based on the findings of this study, it can be concluded that PCR-RFLP is a relatively simple and quick method, completely comparable to the routine methods used for Malassezia identification.