Identification Of Candida Species Research Articles

Identification of Candida species by direct inoculation of cornmeal tween™ 80 agar from blood culture bottles: Rapid and cost-effective approach

Background: This study aimed to compare the accuracy of Candida species identification by direct inoculation of positive blood culture specimens into CornMeal Tween™ (CMT) 80 agar with the conventional methods. Methods: This study was performed in two phases: Phase 1: Inoculation of direct CMT from artificially spiked blood culture bottles with identified Candida species. Phase 2: Direct inoculation of smear-positive bottles on CMT agar for identification. Routine identification method was carried out using urease production, sensitivity to cycloheximide, CMT inoculation, and colony morphology on BiGGY agar. All isolates were also evaluated for sugar assimilation on the biochemical test panel Analytical profile index (API) 20C AUX (BioMerieux). Statistical Analysis Used: The correlation between both methods was determined as percentage agreement rate using kappa score. Results: In the first phase 50 artificially spiked blood culture bottles were included in the study. The sample set consisted of 24% C albicans, 38% C tropicalis, and 22% C glabrata isolates. There was 100% agreement between the directly inoculated and CMT inoculated from colonies at 48 hours. Phase 2: The overall agreement between directly inoculated CMT and conventional method of identification at 72 h was 92%. Conclusion: Direct inoculation of CMT agar can easily differentiate common Candida species microscopically on the basis of chlamydospores, blastopores, and arrangement of pseudo hyphae. However, identification of rare species from specimens containing two different species using direct method is challenging. Therefore, conventional methods including macroscopic/microscopic characteristics and biochemical profile with the aid of API with API 20 AUX would be a better choice for such cases.

Molecular Identification and Antifungal Susceptibility Pattern of Non-albicans Candida Species Isolated from Vulvovaginal Candidiasis

Background:Vulvovaginal candidiasis (VVC) is an important health problem caused by Candida spp. The aim of this study was molecular identification, phylogenetic analysis, and evaluation of antifungal susceptibility of non-albicans Candida isolates from VVC.Methods:Vaginal secretion samples were collected from 550 vaginitis patients at Sayyad Shirazi Medical and Educational Center of Gorgan (Golestan Province, Iran) from May to October 2015. Samples were analyzed using conventional mycological and molecular approaches. Clinical isolates were analyzed with specific PCR using CGL primers, and the internal transcribed spacer region and the D1-D2 domain of the large-subunit rRNA gene were amplified and sequenced. Susceptibility to amphotericin B, fluconazole, itraconazole, and clotrimazole was determined by the guidelines of the Clinical and Laboratory Standard Institute.Results:In total, 35 non-albicans Candida isolates were identified from VVC patients. The isolates included 27 strains of Candida glabrata (77.1%), 5 Candida krusei (Pichia kudriavzevii; 14.3%), 2 Candida kefyr (Kluyveromyces marxianus; 5.7%), and 1 Candida lusitaniae (Clavispora lusitaniae; 2.9%). The fungicides itraconazole and amphotericin B were effective against all species. One isolate of C. glabrata showed resistance to fluconazole and clotrimazole, and 26 isolates of C. glabrata indicated dose-dependent susceptibility to fluconazole. C. lusitaniae was susceptible in a dose-dependent manner to fluconazole and resistant to clotrimazole.Conclusions:Non-albicans Candida spp. are common agents of vulvovaginitis, and C. glabrata is the most common species in the tested patients.

Speciation of Candida using CHROMagar in cases with oral epithelial dysplasia and squamous cell carcinoma.

BackgroundCandida albicans is most frequently isolated from oral cavity but identification of other Candida species such as C. tropicalis, C. krusei, C. glabrata & C. dubliniensis is increasing proportionately. A constant rise in immuno-suppressed patients, widening range of recognized pathogens, and resistance to antifungal drugs are contributing factors which stress the need for species identification of Candida, an opportunistic pathogen. Objectives: 1. To detect the prevalence of Candida albicans and Non albicans Candida albicans (NAC) species in the oral cavity of patients with epithelial dysplasia, Oral squamous cell carcinoma (OSCC) and healthy controls. 2. To identify and differentiate Candidal species using CHROMagar, a differential media.Material and MethodsThe study included smears from 50 patients with histopathological confirmation of epithelial dysplasia & OSCC and 50 normal controls. Candida albicans was identified using Sabouraud dextrose agar media (SDA) as primary culture followed by species identification using CHROMagar on the basis of colony color and morphology.ResultsNon albicans candida predominated (66%) over Candida albicans (34%) in speciation on CHROMagar media in the study group. Non albicans Candida species isolated were C. tropicalis (38%), C. glabrata (24%) and 2 cases showing polyfungal population of C. albicans & C. glabrata.ConclusionsSpecies level isolation of Candida helps in early identification of resistant non Candida strains and prompt treatment of the cases there by preventing the dissemination of infection in case of immuno-compromised individuals. The data presented also supports the use of CHROMagar Candida as a pertinent media for the rapid identification of Candida species directly from clinical specimens in resource challenged settings, which could be helpful in developing appropriate therapeutic strategy and management of patients. Key words:Candida, CHROMagar, epithelial dysplasia, oral cancer.

Identification and antifungal susceptibility of Candida species isolated from the urine of patients in a university hospital in Brazil.

ABSTRACTThe aim of this study was to identify Candida spp. isolated from candiduria episodes at a tertiary hospital in the Midwest region of Brazil, and to determine their susceptibility profiles to antifungal compounds. From May 2011 to April 2012, Candida spp. isolated from 106 adult patients with candiduria admitted to the University Hospital of the Federal University of Mato Grosso do Sul were evaluated. Both, species identification and susceptibility testing with fluconazole-FLC, voriconazole-VRC, and amphotericin B-AmB were carried out using the Vitek 2. To discriminate species of the C. parapsilosis complex, a RAPD-PCR technique using the RPO2 primer was performed. From the total of 106 isolates, 42 (39.6%) C. albicans and 64 (60.4%) Candida non-albicans (CNA) - 33 C. tropicalis, 18 C. glabrata, 5 C. krusei, 4 C. parapsilosis sensu stricto, 2 C. kefyr, 1 C. lusitaniae, and 1 C. guilliermondii were identified. All isolates were susceptible to AmB and VRC, whereas all C. glabrata isolates presented either resistance (5.6%) or dose-dependent susceptibility (94.4%) to FLC. The study of Candida spp. and their resistance profiles may help in tailoring more efficient therapeutic strategies for candiduria.

Merging FT-IR and NGS for simultaneous phenotypic and genotypic identification of pathogenic Candida species.

The rapid and accurate identification of pathogen yeast species is crucial for clinical diagnosis due to the high level of mortality and morbidity induced, even after antifungal therapy. For this purpose, new rapid, high-throughput and reliable identification methods are required. In this work we described a combined approach based on two high-throughput techniques in order to improve the identification of pathogenic yeast strains. Next Generation Sequencing (NGS) of ITS and D1/D2 LSU marker regions together with FTIR spectroscopy were applied to identify 256 strains belonging to Candida genus isolated in nosocomial environments. Multivariate data analysis (MVA) was carried out on NGS and FT-IR data-sets, separately. Strains of Candida albicans, C. parapsilosis, C. glabrata and C. tropicalis, were identified with high-throughput NGS sequencing of ITS and LSU markers and then with FTIR. Inter- and intra-species variability was investigated by consensus principal component analysis (CPCA) which combines high-dimensional data of the two complementary analytical approaches in concatenated PCA blocks normalized to the same weight. The total percentage of correct identification reached around 97.4% for C. albicans and 74% for C. parapsilosis while the other two species showed lower identification rates. Results suggested that the identification success increases with the increasing number of strains actually used in the PLS analysis. The absence of reliable FT-IR libraries in the current scenario is the major limitation in FTIR-based identification of strains, although this metabolomics fingerprint represents a valid and affordable aid to rapid and high-throughput to clinical diagnosis. According to our data, FT-IR libraries should include some tens of certified strains per species, possibly over 50, deriving from diverse sources and collected over an extensive time period. This implies a multidisciplinary effort of specialists working in strain isolation and maintenance, molecular taxonomy, FT-IR technique and chemo-metrics, data management and data basing.

Identification of Candida species isolated from vulvovaginitis using matrix assisted laser desorption ionization-time of flight mass spectrometry.

Background and Purpose:Vulvovaginal candidiasis (VVC) is a common problem in women. The purpose of this study was to identify Candida isolates by matrix assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) from women with vulvovaginitis that were referred to Ghaem Hospital, Mashhad, Iran.Materials and Methods:This study was conducted on 65 clinical samples isolated from women that were referred to Ghaem Hospital. All specimens were identified using phenotyping techniques, such as microscopy and culture on Sabouraud dextrose agar and corn meal agar. In addition, all isolates were processed for MALDI-TOF MS identification.Results:Out of the 65 analyzed isolates, 61 (94%) samples were recognized by MALDI-TOF MS. However, the remaining four isolates (6%) had no reliable identification. According to the results, C. albicans (58.5%) was the most frequently isolated species, followed by C. tropicalis (16.9%), C. glabrata (7.7%), C. parapsilosis (7.7%), and guilliermondii (3.1%).Conclusion:As the findings indicated, MALDI TOF MS was successful in the identification of clinical Candida species. C. albicans was identified as the most common Candida species isolated from the women with VVC. Moreover, C. tropicalis was the most common species among the non-albicans Candida species.

Multicenter Identification and Antifungal Susceptibility Patterns of Candida Species Isolated from Clinical Samples

Background: Invasive fungal infections without proper treatment could lead to high mortality rate, especially in immunocompromised patients. Candida species distribution and drug susceptibility patterns vary in different areas. Understanding the etiologic agents and drug susceptibility patterns in each region are required for the best management of patients with Candida infections. Objectives: The aim of this study was to identify Candida species isolated from clinical samples of six university hospitals in Iran and detect their susceptibility patterns to seven antifungal agents. Methods: Clinical samples from patients with fungal infections were cultured on Sabouraud dextrose agar. Isolated yeasts were identified by API 20C AUX kit, according to the manufacturer’s instructions. Drug susceptibility patterns to amphotericin B, caspofungin, voriconazole, fluconazole, posaconazole, itraconazole and ketoconazole were determined, according to CLSI M27-A3 and S4. Results: In total, 428 species of Candida were isolated from clinical samples (1950 samples). Most isolated species were Candida albicans, followed by C. tropicalis, C. parapsilosis, C. kefyr, C. famata, C. glabrata, C. krusei, C. dubliniensis, C. guilliermondii and C. lusitaniae. Sensitivity rate of C. albicans to amphotericin B, caspofungin, voriconazole, fluconazole, and itraconazole was 96.6, 99.5, 88.6, 90.6, and 52 with MIC90 values equal to 0.25 µg/mL, 0.125µg/mL, 0.125 µg/mL, 2 µg/mL, and 1 µg/mL, respectively. The MIC 90 values for ketoconazole and posaconazole were 0.125µg/mL and 0.064µg/mL, respectively. Different sensitivity to antifungal agents was present in non-albicans Candida species especially in C. krusei, C. glabrata, and C. tropicalis. Conclusions: According to this study, C. albicans is the most prevalent etiologic agent in infected patients and caspofungin is the most effective antifungal agent. Knowledge about etiologic agents and their susceptibility patterns in each region is helpful for successful treatment of the patients. © 2017, Jundishapur Journal of Microbiology.

Diagnosis and management of oral candidosis.

Candida is a fungus (yeast) that is generally regarded as a normal and harmless member of the oral microbiome in humans. Should host defences against these commensals be compromised in any way then Candida can cause clinical signs and symptoms, which manifest as distinct forms of oral candidosis (candidiasis). Candida albicans is the most frequently isolated candidal species from the oral cavity, although a range of non-C. albicans Candida species are being increasingly encountered. The basic principle of the management of candidosis is to identify and eliminate any underlying host predisposing factor. However, in many cases, antifungal therapy will also be required as part of initial management. This article will provide an overview of the isolation, identification and pathogenicity of Candida species encountered within the mouth and relate these to clinical management of oral candidosis.

Candida albicans and Napkin Dermatitis: Relationship and Lesion Severity Correlation

Introduction: Napkin Dermatitis (ND) is a common problem in infancy that affects almost every child during the early months and years of their lifetime. It is a skin disease that becomes a challenge for both parents and physicians because of its frequency and difficulty in eliminating all of the causative factors in diapered infants. Usually Napkin dermatitis is self-limiting but when associated with Candida albicans (C. albicans) seems to be moderate to severe.Aim: The aim of the present study was to determine the colonization of C. albicans in children with Napkin dermatitis and to correlate between intensity of C. albicans colonization and the severity of napkin rash.Patients and Methods: This case-controlled study was conducted at Qassim University pediatric outpatient clinics, during the period from August 2014 to July 2015. Sixty patients with diaper dermatitis and 33 healthy controls were enrolled to this study. Sociodemographic and clinical data were obtained from the parents of each participant using questionnaires Paired (stool and skin) samples were collected from all cases and healthy control children. The samples were cultured on differential and selective chromogenic medium for isolation and initial identification of candida species. Identification confirmation of the isolates was determined by the Vitek 2 compact automated system.Results: Diaper dermatitis shows significant outcome to washing diaper area (per day) (P=0.001), History of diarrhea last 7 Days (P˂0.001), skin lab results (+/-) for Candida albicans, (P˂0.001), skin colony count, (P˂0.001), However, there is no correlation to age (P=0.828), gender (P=0.368) and feeding style (P=0.401).Conclusion: The severity score of napkin dermatitis was significantly observed among cases with diaper dermatitis (p-value<0.001) and control children (p-value<0.001) respectively.Keywords: Candida albicans; Napkin dermatitis; Diaper dermatitis; Vitek 2 compact system; Qassim.

Comparison of CHROMagar, polymerase chain reaction-restriction fragment length polymorphism, and polymerase chain reaction-fragment size for the identification of Candida species.

Background and Purpose:The epidemiological alteration in the distribution of Candida species, as well as the significantly increasing trend of either intrinsic or acquired resistance of some of these fungi highlights the need for a reliable method for the identification of the species. Polymerase chain reaction (PCR) is one of the methods facilitating the quick and precise identification of Candida species. The aim of this study was to compare the efficiency of CHROMagar, PCR-restriction fragment length polymorphism (PCR-RFLP), and PCR-fragment size polymorphism (PCR-FSP) assays in the identification of Candida species to determine the benefits and limitations of these methods.Materials and Methods:This study was conducted on 107 Candida strains, including 20 standard strains and 87 clinical isolates. The identification of the isolates was accomplished by using CHROMagar as a conventional method. The PCR-RFLP assay was performed on the entire internal transcribed spacer (ITS) region of ribosomal DNA (rDNA), and the consequent enzymatic digestion was compared with PCR-FSP results in which ITS1 and ITS2 regions were separately PCR amplified. In both molecular assays, yeast identification was carried out through the specific electrophoretic profiles of the PCR products.Results:According to the results, the utilization of CHROMagar resulted in the identification of 29 (33.3%) Candida isolates, while the PCR-RFLP and PCR-FSP facilitated the identification of 83 (95.4%) and 80 (91.9%) clinical isolates, respectively. The obtained concordances between CHROMagar and PCR-RFLP, between CHROMagar and PCR-FSP, as well as between PCR-RFLP and PCR-FSP were 0.23, 0.20, and 0.77, respectively.Conclusion:The recognition of the benefits and limitations of PCR methods allows for the selection of the most efficient technique for a fast and correct differentiation. The PCR-RFLP and PCR-FSP assays had satisfactory concordance. The PCR-FSP provides a rapid, technically simple, and cost-effective method for the identification of Candida species. Nevertheless, to accurately differentiate among the taxonomically related species, PCR-RFLP should be implemented.

Conventional and Molecular Identification of Pathogenic and Keratinophilic Fungi Isolated from Feathers of Bird in Erbil City

During the period of August 2014 to January 2015, two hundred twenty four samples from 43 species of bird pets were collected from feathers then examined. The results of isolated feathers samples shows that Chaetomium species constituted the majority of saprophytic fungi (27.5%) followed by Aspergillus spp. (22.8%), Humicola spp. (8.7%), Emericella spp. (8.7%), Penicillium spp. (8.6%), Stachybotrys spp. (4.5%), Acremonium spp. (3.7%), Rhizopus spp. (3.6%), Drechslera spp. (3.1%), Cunninghamella spp. (2.4%), Mucor spp. (2.4%), Fusarium spp. (1.8%), F. dimerum (0.9%) and Sepidonium spp. (1.4%). The highest rate of isolated dermatopytes was Trichophyton which was (75%) followed by Arthroderma spp. (17.3%), Microsporum canis (5.1%) and M. gypsium (2.6%). Among keratinophilic fungi Chrysosporium spp. has the highest rate of isolated fungi by (91.4%) followed by Fonsecaea spp. (2.8%), Scopulariopsis spp. (2.8%), S. brevicaulis (0.9%) and Syncephalastrum spp. (2.1%). Among yeasts Candida spp . was the highest which was (74.2%) followed by Trichosporonasahii (13.8%), Geotricumklebahnii (5.8%), G. candidum (2.4%), Cryptococcus laurentii (3.8%). Genomic DNA was extracted from common highly dermatophytes, keratinophilic and Candida spp. by using EZ-10 Spin Column Fungal Genomic DNA Mini-Preps Kit, with high purity 1.7-1.8, the ITS regions from all extracted DNA were amplified by using ITS1/ITS4 primer set, for identification of dermatophytes, Keratinophilic and Candida species. The products were electrophoresed through 2% agarose gels in TBE buffer and visualized by staining with ethidium bromide under UV irradiation.

IDENTIFICATION OF CANDIDA SPECIES ON THE SKIN OF DIABETES MELLITUS PATIENTS

Background: Intertriginous candidiasis is a type of candidiasis which the lesion lies in the folds of the armpit, groin, intergluteal, folding of the breast, between the fingers or the toes, glans penis, and the umbilicus. One of the risk factors for intertriginous candidiasis is diabetes mellitus (DM). DM is a typical clinical syndrome characterized by the presence of hyperglycemia which is caused by deficiency or the decrease of insulin effectivity. Fungal infection of the skin with Candida species becomes easier in patients with immunocompromised conditions such as DM patients.Objective: To identify the Candida species in DM patient's skin.Method: This research is an observational descriptive study with cross sectional design. This research identifies Candida species from 30 DM patients at The Public Health Center Sentosa BaruMedan who met the inclusion and exclusion criteria by skin scraping with KOH 10% and slide culture.RESULTS: The location of the rash was found in the crotch folds 63.3% and in the intergluteal 36.7%. All samples (100%) showed positive fungus structure with KOH 10% examination. On theexamination of slide culture with Saboraoud Dextrose Agar media and identification of slide colony with Corn Meal Agar medium as many as 60% Candida spesies was found with overallspecies are Candida albicans.Conclusions: The identification of Candida Species on the skin of DM patients showed as many as 60% Candida albicans was found

Antifungal Susceptibility of Candida Species Isolated from Cancer Patients with Oral Lesions Undergoing Chemotherapy

Background: Oral candidiasis is the most common opportunistic infection of the oral cavity in patients undergoing chemotherapy. Identification of Candida species and the corresponding susceptibility to antifungal agents can be helpful in the management of cancer patients. Objectives: The purpose of this study was to determine the susceptibility patterns of Candida species against 3 antifungal agents and define clinical practice guidelines for the prevention and treatment of oral candidiasis in cancer patients. Methods: A total of 12 positive samples of oral lesions caused by Candida species were isolated from patients undergoing chemotherapy through direct examination and culture on CHROMagar Candida medium. Stock cultures were grown on sabouraud dextrose agar and DNA extracts. Then, polymerase chain reaction (PCR) was performed and the products were sequenced. The microdilution method was applied at different concentrations of fluconazole, amphotericin B, and nystatin. The minimum inhibitory concentration (MIC) and minimum fungicidal concentration (MFC) of each species were compared. Results: The species distribution of Candida isolates was as follows: C. albicans, 6 (50%); C. krusei, 3 (25%); and C. tropicalis, 3 (25%). The male-to-female ratio was 8:4, and the mean age of cancer patients was 51.25 years (range, 25-81 years). Overall, 100% and 83% of C. albicans were resistant to nystatin and fluconazole, respectively. All Candida species showed the lowest and highest resistance to amphotericin B (8.3%) and nystatin (66.7%), respectively. Conclusions: DNA sequencing showed that C. albicans is the most commonly identified species in the oral cavity of cancer patients. Amphotericin B, compared to fluconazole and nystatin, is a more suitable antifungal drug for oral candidiasis. Oral hygiene involves dental cleaning, and management of poor denture hygiene and xerostomia can be helpful in eliminating Candida species in patients undergoing chemotherapy.

Identification of Candida species in vaginal flora using conventional and molecular methods

Vaginal swab samples were collected both from women with signs of acute vulvovaginal candidiasis (VVC; n=270) and from healthy controls (n=100). The microscopic examination revealed a significant difference in the proportion of positive cultures of Candida between the VVC (33%) and control (21%) groups. To determine the frequency of different species, positive cultures were analyzed using the germ tube test, CHROMagar medium, API 20 AUX System and DNA analysis. CHROMagar and API 20 AUX System tests identified 67% of samples as C.albicans. Out of these, 42% appeared to be C.dubliniensis when the specific primers in the polymerase chain reaction (PCR) were used. We observed the prevalence of Candida species isolated from the vagina in descending order as follows: C.albicans (38-39%), C.glabrata (32-33%), C.dubliniensis (28-29%) and C.tropicalis (0-1%) for both the VVC and the control group.

Epidemiologic Features of Vulvovaginal Candidiasis in Women of Childbearing Age in a Combined Military Hospital

Introduction: Vulvovaginal Candidiasis is a reproductive tract infection which remains a common cause of morbidity adversely affecting women’s physical and emotional health. Vaginal discharge in the childbearing age group is the most common complaint encountered every day both by gynaecologists and general practitioners. Among the cases of symptomatic vaginal discharge, Bacterial vaginosis is the commonest cause followed by Candidiasis and Trichomoniasis. It is characterized by curd-like vaginal discharge and itching and is associated with considerable health and economic costs.
 Objective: To detect the prevalence of Candida species in patients with complaints of vaginal discharge of reproductive age group attending at Child Welfare Centre (CWC), Gynaecology Outpatient Department (GOPD) and Medical Inspection Room (MI Room) of Combined Military Hospital (CMH) Ghatail.
 Materials and Methods: A cross-sectional descriptive study was conducted at CMH Ghatail during the period of April 2017 to October 2017. A total of 183 patients of reproductive age group with the complaints of vaginal discharge attending CWC, GOPD and MI Room of CMH Ghatail were included in this study. Two specimens of High Vaginal Swabs (HVS) were collected from each patient, one for direct microscopy and another for culture. For every patient, two wet film preparations and two Gram’s staining were done. Specimens were cultured on Blood Agar Medium and Sabourad’s Dextrose Agar Medium. Identification of Candida species was carried out by Wet film, Gram’s staining and culture examination.
 Results: According to the results of this study, the prevalence of Vulvovaginal Candidiasis (VVC) was found in 57(31.15%) of the cases. The maximum number of patients 126(66.66%) were found between 26 years to 35 years of age group. On the other hand, amongst 57 VVC positive cases, 44(77.19%)were identified as Candida albicans and 13(22.81%) were Candida non-albicans.
 Conclusion: Candida albicans is the predominant organism amongst candida species identified from VVC in this study. The high prevalence necessitates adequate screening of woman with vaginal discharge in order to give appropriate treatment and to avoid complications associated with it.
 Journal of Armed Forces Medical College Bangladesh Vol.13(1) 2017: 13-16

Automated Identification and Antifungal Susceptibility Testing of Candida Species Using Vitek 2 Compact System in ICUs and Pediatric Oncology Unit, Alexandria, Egypt

Background: The epidemiology of candida species has changed over the last two decades. Early species identification and rapid antifungal susceptibility testing (AFST) is mandatory in such critical patients. Objectives: To determine the prevalence of different types of Candida species in ICU and pediatric oncology patients as well as their antifungal susceptibility profile using the Vitek 2 compact system. Methodology: 1023 candida isolates were collected from different clinical samples between May 2014 and February 2017. Identification and antifungal susceptibility testing were performed using Vitek 2 compact system. Results: In the present study fifteen candida species were identified. Non albicans Candida species (NAC) were predominant (65.6%). Candida albicans was the most prevalent species (34.5%). Candida tropicalis (c.tropicalis) was the most common isolated NAC species and the second common isolated species (32.8%), followed by Candida glabrata (C.glabrata) (17.3%), Candida Parapsilosis (C.parapsilosis)(7.5%), accounting for 92.1% of the total number of isolates. Uncommon Candida species were also isolated in this study.C. tropicalis was the most frequent species isolated from urine (50.9%), while C. albicans was the most frequent species isolated (43.3%) & (30.8%) from respiratory and blood samplesrespectively.C. parapsilosis was the most common species isolated from indwelling devices (44.4%). Regarding antifungal susceptibility profile (5.1%) and (2 %) of C.albicans were resistant to fluconazole, and voriconazole repectively. C. tropicalis (24.6%) and (17.5%) were resistant to Fluconazole and voriconazole respectively. (13.9%), (7.2%), (2.4%) of C.glabrata isolates were resistant to fluconazole, voriconazole and amphotericin respectively. All C. parapsilosis isolates were sensitive to fluconazole, voriconazole. Although resistant to fluconazole, all C. krusei isolates were susceptible to voriconazole, while, (84.6%) of isolates were resistant to flucytosine. Conclusions: The increased isolation rates of NAC species and gradual shift in the antifungal susceptibility profile underlines the need of early and accurate diagnosis of infecting Candidaspecies along antifungal susceptibility testing.

A new assay based on terminal restriction fragment length polymorphism of homocitrate synthase gene fragments for Candida species identification

Candida sp. have been responsible for an increasing number of infections, especially in patients with immunodeficiency. Species-specific differentiation of Candida sp. is difficult in routine diagnosis. This identification can have a highly significant association in therapy and prophylaxis. This work has shown a new application of the terminal restriction fragment length polymorphism (t-RFLP) method in the molecular identification of six species of Candida, which are the most common causes of fungal infections. Specific for fungi homocitrate synthase gene was chosen as a molecular target for amplification. The use of three restriction enzymes, DraI, RsaI, and BglII, for amplicon digestion can generate species-specific fluorescence labeled DNA fragment profiles, which can be used to determine the diagnostic algorithm. The designed method can be a cost-efficient high-throughput molecular technique for the identification of six clinically important Candida species.

Evaluation of Chromogenic agar media for rapid identification of Candida species.

Rapid identification of Candida isolates to the species level is essential in order to optimize the antifungal treatment. This study aimed to isolate and identify different species of Candida from various clinical specimens and to evaluate the use of chromogenic agar media as primary culture media for culture of Candida as well as for rapid identification of Candida species. A total of 100 different clinical specimens were studied (oral swab 35, high vaginal swab 28, catheterized urine 15, nail 14, bronchoalveolar lavage 04 and peritoneal fluid 04). Isolation of Candida species was done by primary culture in Sabouraud Dextrose Agar (SDA). Subsequent identification of species was performed by germ tube test, carbohydrate assimilation test (with commonly used twelve sugars) and subculture in chromogenic agar medium. Out of 64 isolated Candida, C. albicans 33 (51.53%) was the most predominant Candida species followed by C. tropicalis 17 (26.56%). The species of C. glabrata was 4 (6.25%), C. parapsilosis 4 (6.25%), C. krusei 3 (4.68%) and C.guilliermondii 2 (3.2%). One of the isolated Candida species was unidentified. The sensitivity and specificity of chromogenic agar media for C. albicans were as 96.97% and 96.87% respectively. The sensitivity and specificity for C. tropicalis were 94.12% and 97.87% respectively. C. krusei and C. glabrata both showed 100% sensitivity and specificity on chromogenic agar media. Efficacy of chromogenic agar media is nearly similar to carbohydrate assimilation method in species identification of Candida.Bangladesh J Med Microbiol 2015; 9 (1): 22-26

Congenital Cutaneous Candidiasis: Prompt Systemic Treatment Is Associated With Improved Outcomes in Neonates.

Congenital cutaneous candidiasis (CCC) is a challenging diagnosis due to various rash presentations. Inadequate early treatment is associated with high rates of dissemination and death. The effects of early diagnosis, dermatologic presentation, and antifungal treatment on outcomes are lacking. CCC cases were reviewed from 2 academic neonatal intensive care units (NICUs) from 2004 to 2015. We defined CCC as a diffuse rash involving the body, extremities, face or scalp, and/or funisitis, presenting in the first week (≤7 days), with identification of Candida species from skin or mucous membrane cultures, and/or by culture or staining of the placenta or umbilical cord. CCC occurred in 0.1% of all NICU admissions (21 of 19 303) and 0.6% of infants <1000 grams birth weight. Median gestational age of CCC infants was 26 3/7 (range, 23 0/7-40 4/7) weeks. Skin findings were commonly present on the day of birth [median (range): 0 (0-6) days], appearing most frequently as a desquamating, maculopapular, papulopustular, and/or erythematous diffuse rash. When systemic antifungal therapy was started empirically at the time of rash presentation and continued for a median (interquartile range) of 14 (14-15) days, all patients survived and none developed dissemination. Delaying systemic treatment, exclusive use of nystatin, and treating for <10 days was associated with Candida bloodstream dissemination. CCC is an invasive infection that presents as a diffuse rash in preterm and term infants. Prompt systemic antifungal treatment at the time of skin presentation for ≥14 days prevents dissemination and Candida-related mortality.

Development of a real-time PCR assay for the direct detection of Candida species causing Vulvovaginal candidiasis

Identification of Candida species by traditional methods can be time-consuming and have limited analytical sensitivity. We developed a multiplex real-time PCR assay for detection and differentiation of Candida species causing vulvovaginal candidiasis (VVC). Overall, this PCR assay is a powerful diagnostic tool offering superior accuracy, sensitivity, and specificity.