Rapid Identification Of Candida Species Research Articles

An Integrated and Multi-Target Nucleic Acid Isothermal Analysis System for Rapid Diagnosis of Vulvovaginal Candidiasis.

Rapid identification of Candida species is significant for the diagnosis of vulvovaginal candidiasis (VVC). An integrated and multi-target system for the rapid, high-specificity, and high-sensitivity detection of four Candida species was developed. The system consists of a rapid sample processing cassette and a rapid nucleic acid analysis device. The cassette could process the Candida species to release nucleic acids in 15 min. The released nucleic acids were analyzed by the device as fast as within 30 min, using the loop-mediated isothermal amplification method. The four Candida species could be simultaneously identified, with each reaction using only 1.41 µL of reaction mixture, which was low cost. The RPT (rapid sample processing and testing) system could detect the four Candida species with high sensitivity (<2 CFU/reaction) and high specificity. The system also processed and analyzed 32 clinical samples, giving the results with high clinical sensitivity and specificity. Hence, the system was a significant and effective platform for the diagnosis of VVC. Furthermore, the period of validity of the reagents and chips used in the system was >90 days, and the system could also be used for the detection of bacteria.

Non Candida albicans Candida (NCAC) of oral cavity in head and neck cancer patients under cancer therapy: Prevalance, species identification and antimycotic sensitivity pattern

Introduction: Cancer patients remain at risk for developing serious infections due to oropharyngeal Candida colonisation as the incidence of Candidiasis continues to rise after chemotherapy (CT), Radiochemotherapy (RCT) or Radiotherapy (RT) for head and neck cancer. So, accurate and rapid identification of candida species is very important in clinical laboratory. Objective: To study the prevalance, species identification and antimycotic sensitivity pattern of Candida spp. of oral cavity in patients of head & neck cancer undergoing chemotherapy (CT) & Radiochemotherapy (RCT) and compare with control group. Materials and Methods: This is prospective cross sectional and case control analysis including 110 patient in study group; chemotherapy (CT)-55 patient, Radiochemotherapy (RCT)-55 patients and 50 healthy individuals as control. Candida isolates identified by standrad methods using morphological, biochemical & chromogenic property of different Candida spp. and antimycotic sensitivity applied by disc diffusion method. Results: Prevalence of Candida spp. colonization seen in 50% patients of study and 20% in control group (p value- 0.004) ; patients on RCT (63.6%) have higher prevelance as compare to CT (36.4%), (p 0.007). C. tropicalis was most common (38%) species followed by C. albicans (11%) & C. glabrata (11%); most of isolates were sensitive to Amphotericin B and Nystatin and least sensitive to Fluconazole. Conclusion: High prevalence of Candidal carriage and the species variation and changing pattern of antifungal susceptibility of Candida spp in patients undergoing chemotherapy and radiation therapy require isolation and speciation of the causative Candida sp. and prophylaxis is needed to prevent infection among these patients. Keywords: Candida species, Non Candida albicans Candida (NCAC), Oral cavity, Head & neck cancer, Radiochemotherapy.

Molecular diagnosis and antifungal resistance and biofilm production profiles among Candida species isolated from immunocompromised patients at Menoufia University Hospitals

Background: Accurate and rapid identification of Candida species is necessary for proper diagnosis and treatment of candidiasis due to emergences of drug-resistant strains especially among immunocompromised patients. Objectives: Identification of Candida clinical isolates to the species level using different phenotypic and molecular methods. Biofilm-forming ability and antifungal resistance were also studied. Methodology: Sixty-nine Candida strains were isolated from 220 immunocompromised patients. Identification was performed using chromogenic Candida agar, VITEK 2 system and multiplex polymerase chain reaction (PCR). Biofilm formation was detected by the tube method and antifungal susceptibility was tested using the VITEK2 system. Results: The most common source of Candida isolates was from urine (33.3%) and ICUs (56.6%). VITEK 2 system detected 9 spp.: C. albicans (34.8%), C. tropicalis (21.7%), C. famata (8.7%), C. lusitaniae (7.2%), C. cruzi (7.2%), C. ciferri (5.8%), C. dubliniensis (5.8%), C. parapsilosis (5.8 %) and C. glabrata. Candida isolates showed high resistance to flucytocine (49.3%), and high sensitivity to fluconazole, micafungin, voriconazole and caspofungin (88.4%, 81.2% and 81.2 % respectively). Only 30.4% of all Candida isolates were biofilm producers. There was a positive relationship between antifungal resistance and biofilm formation among Candida isolates. Conclusion: C. albicans was the predominant species. Chromogenic Candida agar and VITEK 2 system were valuable tests compared to PCR in speciation of Candida isolates. Antifungal susceptibility was significantly related to biofilm production and its evaluation is important for proper treatment..

Kan Kültürlerinde Tespit Edilen Candida İzolatlarının Dağılımı ve Antifungal Duyarlılıkları

Candidemia is a serious clinical condition that can result in mortality. This condition is particularly common in intensive care patients. This retrospective study aimed to identify Candida strains isolated from blood cultures and to determine antifungal susceptibility patterns. For this purpose, isolated Candida species were identified and, their susceptibility to amphotericin B, caspofungin, flucytosine, fluconazole, micafungin and voriconazole were investigated. VITEK 2 automated system was used to identify and detect antifungal susceptibility of Candida species that were subjected to Gram staining and germ tube tests on Candida species detected from blood cultures sent to our laboratory between January 2014 and December 2018. Candida spp. isolation rate was found most frequently (n=157; 64.9 %) in intensive care units; followed by internal clinics (n=64; 26.5 %) and surgical clinics (n=21; 8.6 %). The most frequently isolated species among these Candida isolates are Candida albicans (100/242; 41.3 %) and Candida parapsilosis (92/242; 38 %). Especially in intensive care patients, rapid identification of Candida species and determination of their antifungal susceptibilities are important for planning treatment. It is thought that collecting data reflecting regional resistance status at certain intervals will be guiding in terms of treatment approaches.

Comparison of methods for detection of Candida in bronchoalveolar lavage (BAL) of cancer patients

An accurate and rapid identification of Candida species in cancer patients with pulmonary symptoms can provide important information for effective treatment. Candida infections represent an increasing cause of morbidity and mortality in patients receiving immunosuppressive chemotherapy for cancer, organ transplantation or in immunocompromised. We used conventional methods such as culture, fermentation reactions, morphology and molecular methods based on the ribosomal DNA repetitive regions or the Internal Transcribed Spacer (ITS) for the identification of Candida species. Seventy bronchial specimens of Bronchoalveolar Lavage (BAL) from cancer patients at Shaukat Khanum Memorial Cancer Hospital and Research Center. Lahore, Pakistan were included in this study. Seventy cancer patients were diagnosed on the basis of histological profile. Candida detected by conventional methods using Sabouraud Dextrose Agar (SDA), potassium hydroxide (KOH) preparation, germ tube test, fermentation reactions and Gomori methanamine-silver stain (GMS). Thirty (42 %) positive isolates of Candida species were obtained by culture, twenty (28 %) isolates were germ tube test positive while thirty isolates (42 %) were positive by PCR method. In conclusion, the results of our study showed that the PCR based detection methods are significantly better and can detect Candida with more accuracy and specificity as compared to conventional methods. Our study would pave the path for optimization of protocols for detection of Candida in cancer patients.

Rapid species identification of Candida directly from blood culture broths by Sepsityper-MALDI-TOF mass spectrometry: impact on antifungal therapy.

We applied Sepsityper matrix-assisted laser desorption ionisation time of flight mass spectrometry (MALDI-TOF-MS) technology on positive blood culture broths for rapid species identification. Of the 74 patients with candidaemia, 25 had the species identified on the day of the positive blood culture directly from the broth (rapid identification group) while the remaining 49 had the species identified from culture (conventional identification group). Three (13.6%) out of 22 treated patients in the rapid identification group received echinocandin compared to 20/45 (44.4%) in the conventional identification group. The appropriateness of therapy was 90.9% in the rapid identification group and 62.2% in the conventional identification group (p = 0.01). Cost savings were more than £10,000 in the first three days of treatment. Sepsityper-MALDI-TOF-MS is a useful tool in supporting antifungal stewardship programmes.

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.

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

Use of cost effective and rapid molecular tools for identification of Candida species, opportunistic pathogens.

Background and Purpose :Candidiasis is a widespread fungal infection caused by different Candida species. Rapid identification of Candida species in clinical laboratory is becoming increasingly important since the identification and discrimination of ethological agents for early treatment. We aimed at molecular identification of commonly Candida species isolated from clinical samples by using both PCR-RFLP assay and amplification of hwp1 gene. Materials and Methods:Clinical samples comprising of vaginal specimens ,cutaneous, sputum, bronchoalveolar lavage(BAL,( and blood cultures were recovered from suspected patients. Candida isolates were initially identified phenotypically and confirmed by molecular approaches based on restriction fragment length polymorphism (PCR-RFLP (with MspI restriction enzyme. Amplification of hwp1 gene was performed for discrimination of C. albicans from C. dubliniensis and C. africana. Results:The most abundant species were C. albicans (n=67; 44.6 %), C. glabrata (n=10; 20 %), C. tropicalis (n=20; 13.3 %), C. krusei (n=12; 8 %), C. parapsilosis (n=11; 7.3 %). Out of 67 C. albicans species, 6 species identified as C. dubliniensis and 4 species identified as C. africana. Conclusion:High frequency of non-albicans Candida species and differences in levels of susceptibility to the antifungal agents are important issues in medicine .Therefore, to manage the Candida-related infections properly, molecular diagnostic methods would be fast, reliable and even cost-effective approaches for identification of Candida species.

CHROMagar as a primary isolation medium for rapid identification of Candida and its role in mixed Candida infection in sputum samples

Introduction: Candida species occurs as commensal organisms but in immunocompromised patients and those on long term antibiotics, these unrecognized opportunistic fungi may become fatal. Although C. albicans is the most frequently isolated pathogen, the incidence of infections due to non-albicans Candida are on the increase especially with fluconazole resistant strains. Identification of Candida to species level therefore becomes important for selecting the appropriate antifungal agents. The need for rapid identification and the difficulty in detecting mixed cultures on the traditional Sabouraud’s dextrose agar (SDA) have led to the development of chromogenic media. Aim: This study was done to evaluate the usefulness of CHROMagar Candida for rapid identification of Candida species and to identify mixed Candida infection from sputum samples. Materials and methods: A total of 126 sputum samples were inoculated on CHROMagar Candida. The plates were incubated at 37?C for 48 hrs and the growth identified based on the colour of the colony. Results: Single yeast infection was observed in 113 samples (89.7%) and 11 samples showed mixed infection (8.7%). In our study, C.albicans (61.5%) was the predominant species and among the non-albicans Candida, C. tropicalis (18.5%) was the commonest species. Conclusion: Certainly CHROMagar Candida will be useful as a primary culture medium for identifying the yeast infections directly from the clinical specimens and also for the detection of mixed yeast infections which cannot be done on conventional SDA. Hence it helps us in selecting the appropriate antifungal agents for therapy and prophylaxis. Keywords: Candida albicans, Non-albicans Candida, CHROMagar, Mixed yeast infections, Antifungal agents

Identification of Candida Species Using MP65 Gene and Evaluation of the Candida albicans MP65 Gene Expression in BALB/C Mice.

Background:Systemic candidiasis is a major public health concern. In particular, in immunocompromised people, such as patients with neutropenia, patients with Acquired Immune Deficiency Syndrome (AIDS) and cancer who are undergoing antiballistic chemotherapy or bone marrow transplants, and people with diabetes. Since the clinical signs and symptoms are nonspecific, early diagnosis is often difficult. The 65-kDa mannoprotein (MP65) gene of Candida albicans is appropriate for detection and identification of systemic candidiasis. This gene encodes a putative b-glucanase mannoprotein of 65 kDa, which plays a major role in the host-fungus relationship, morphogenesis and pathogenicity.Objectives:The current study aimed to identify different species of Candida (C. albicans, C. glabrata and C. parapsilosis) using the Polymerase Chain Reaction (PCR) technique and also to evaluate C. albicans MP65 gene expression in BALB/C mice.Materials and Methods:All yeast isolates were identified on cornmeal agar supplemented with tween-80, germ tube formation in serum, and assimilation of carbon sources in the API 20 C AUX yeast identification system. Polymerase Chain Reaction was performed on all samples using species-specific primers for the MP65 65 kDa gene. After RNA extraction, cDNA synthesis was performed by the Maxime RT Pre Mix kit. Candida albicans MP65 gene expression was evaluated by quantitative Real-Time (q Real-Time) and Real-Time (RT) PCR techniques. The 2-ΔΔCT method was used to analyze relative changes in gene expression of MP65. For statistical analysis, nonparametric Wilcoxon test was applied using the SPSS version 16 software.Results:Using biochemical methods, one hundred, six and one isolates of clinical samples were determined as C. albicans, C. glabrata and C. parapsilosis, respectively. Species-specific primers for PCR experiments were applied to clinical specimens, and in all cases a single expected band for C. albicans, C. glabrata and C. parapsilosis was obtained (475, 361 and 124 base pairs, respectively). All species isolated by culture methods (100% positivity) were evaluated with PCR using species-specific primers to identify Candida species. Relative expression of Mp65 genes increased significantly after C. albicans injection into the mice (P < 0.05).Conclusions:The results of the current study showed that the PCR method is reproducible for rapid identification of Candida species with specific primers. Mp65 gene expression of C. albicans after injection into the mice was 2.3 folds higher than before injection, with this difference being significant. These results indicated that increase of Mp65 gene expression might be an early stage of infection; however definitive conclusions require further studies.

Candida identification: a journey from conventional to molecular methods in medical mycology

The incidence of Candida infections have increased substantially in recent years due to aggressive use of immunosuppressants among patients. Use of broad-spectrum antibiotics and intravascular catheters in the intensive care unit have also attributed with high risks of candidiasis among immunocompromised patients. Among Candida species, C. albicans accounts for the majority of superficial and systemic infections, usually associated with high morbidity and mortality often caused due to increase in antimicrobial resistance and restricted number of antifungal drugs. Therefore, early detection of candidemia and correct identification of Candida species are indispensable pre-requisites for appropriate therapeutic intervention. Since blood culture based methods lack sensitivity, and species-specific identification by conventional method is time-consuming and often leads to misdiagnosis within closely related species, hence, molecular methods may provide alternative for accurate and rapid identification of Candida species. Although, several molecular approaches have been developed for accurate identification of Candida species but the internal transcribed spacer 1 and 2 (ITS1 and ITS2) regions of the rRNA gene are being used extensively in a variety of formats. Of note, ITS sequencing and PCR-RFLP analysis of ITS region seems to be promising as a rapid, easy, and cost-effective method for identification of Candida species. Here, we review a number of existing techniques ranging from conventional to molecular approaches currently in use for the identification of Candida species. Further, advantages and limitations of these methods are also discussed with respect to their discriminatory power, reproducibility, and ease of performance.

English

Traditional methods that are used to identify clinical isolates of&nbsp;Candida&nbsp;species are time-consuming and not appropriate for rapid, accurate and reliable identification. The aim of this study was to characterize&nbsp;Candida&nbsp;species isolated from fingernail onychomycosis by using polymerase chain reaction (PCR) followed by restriction fragment length polymorphism (RFLP) with three different restriction enzymes HaeIII, DdeI and BfaI. 16Candida&nbsp;isolates recovered from infected fingernail samples were selected and phenotypically identified by API 20C identification kit. Also, the five species were identified genotypically with PCR-RFLP analysis. 16 clinical&nbsp;Candida&nbsp;isolates comprising five species were included in this study; three&nbsp;Candida&nbsp;species were identified using conventional API 20C:&nbsp;C. albicans&nbsp;(9/16),&nbsp;C. parapsilosis&nbsp;(3/16) and&nbsp;C. glabrata&nbsp;(1/16) with three unidentified isolates. Five species were identified based on genotypic identification with PCR-RFLP analysis revealing&nbsp;C. albicans (9/16), C. parapsilosis&nbsp;(4/16),C. glabrata,&nbsp;C. guilliermondii&nbsp;and&nbsp;C. tropicalis&nbsp;(1/16 for each). It was concluded that the PCR-RFLP analysis of rDNA enabled more easy, rapid and precise identification ofCandida&nbsp;species compared to phenotypic methods. RFLP analysis demonstrated that HaeIII is the most differentiating enzyme that distinguished&nbsp;C. albicans&nbsp;from non&nbsp;C. albicans&nbsp;species. BfaI maintained the differentiation of these non&nbsp;C. albicans&nbsp;species and DdeI digestion confirmed&nbsp;C. albicans&nbsp;species. &nbsp; Key words:&nbsp;Onychomycosis,&nbsp;Candida&nbsp;species, polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP).

Identification of Candida species in the clinical laboratory: a review of conventional, commercial, and molecular techniques

In healthy individuals, Candida species are considered commensal yeasts of the oral cavity. However, these microorganisms can also act as opportunist pathogens, particularly the so-called non-albicans Candida species that are increasingly recognized as important agents of human infection. Several surveys have documented increased rates of C. glabrata, C. tropicalis, C. guilliermondii, C. dubliniensis, C. parapsilosis, and C. krusei in local and systemic fungal infections. Some of these species are resistant to antifungal agents. Consequently, rapid and correct identification of species can play an important role in the management of candidiasis. Conventional methods for identification of Candida species are based on morphological and physiological attributes. However, accurate identification of all isolates from clinical samples is often complex and time-consuming. Hence, several manual and automated rapid commercial systems for identifying these organisms have been developed, some of which may have significant sensitivity issues. To overcome these limitations, newer molecular typing techniques have been developed that allow accurate and rapid identification of Candida species. This study reviewed the current state of identification methods for yeasts, particularly Candida species.

A real-time PCR assay for the differentiation of Candida species

We established a real-time PCR assay for the detection and strain identification of Candida species and demonstrated the ability to differentiate between Candida albicans the most common species, and also Candida parapsilosis, Candida glabrata, Candida tropicalis and Candida dubliniensis by LightCycler PCR and melting curve analysis. The DNA isolation from cultures and serum was established using the QIAmp Tissue Kit. The sensitivity of the assay was ≥ 2 genome equivalents/assay. It was possible to differentiate all investigated Candida species by melting curve analysis, and no cross-reaction to human DNA or Aspergillus species could be observed. The established real-time PCR assay is a useful tool for the rapid identification of Candida species and a base technology for more complex PCR assays. We carried out initial steps in validation of a PCR assay for the detection and differentiation of medically relevant Candida species. The PCR was improved by generating PCR standards, additional generation of melting curves for species identification and the possibility to investigate different specimens simultaneously.

Use of CHROMagar Candida for the presumptive identification of Candida species directly from clinical specimens in resource-limited settings

Introduction: Identification of yeast isolated from clinical specimens to the species level has become increasingly important. Ever-increasing numbers of immuno-suppressed patients, a widening range of recognized pathogens, and the discovery of resistance to antifungal drugs are contributing factors to this necessity.Material and methods: A total of 487 yeast strains were studied for the primary isolation and presumptive identification, directly from clinical specimen. Efficacy of CHROMagar Candida has been evaluated with conventional methods including morphology on Corn meal–tween 80 agar and biochemical methods by using API 20 C AUX.Results: The result of this study shows that CHROMagar Candida can easily identify three species of Candida on the basis of colonial color and morphology, and accurately differentiate between them i.e. Candida albicans, Candida tropicalis, and Candida krusei. The specificity and sensitivity of CHROMagar Candida for C. albicans calculated as 99%, for C. tropicalis calculated as 98%, and C. krusei it is 100%.Conclusion: The data presented supports the use of CHROMagar Candida for the rapid identification of Candida species directly from clinical specimens in resource-limited settings, which could be very helpful in developing appropriate therapeutic strategy and management of patients.

Hichrom candida agar for identification of candida species

Chromogenic media are frequently used in direct and rapid identification of yeasts because different Candida species produce unique colors on these media. We used 60 isolates of Candida species including 30 C. albicans, 10 C. parapsilosis, 11 C. glabrata, five C. tropicalis, and four C. dubliniensis, isolated from various clinical specimens, to evaluate the performance of HiChrome Candida agar. These strains had been identified by germ tube test, morphology on cornmeal agar, chlamydospore formation on tobacco agar and sugar assimilation tests. The sensitivity and specificity results were: C. albicans (96.55 and 96.42%); C. parapsilosis (80 and 98.03%), C. glabrata (90.90 and 88.23%), C. tropicalis (100 and 100%) and C. dubliniensis (60 and 96.55%) respectively. HiChrom Candida agaris medium has been useful and capable of presumptive, rapid identification of Candida species within 48 hours.

Early diagnosis of blood culture isolates in patients with candidemia using PNA FISH

Candidemia is associated with high mortality, especially with neutropenic and intensive care unit patients. A delay in early, effective antifungal therapy has been associated with increased mortality and hospital costs. Peptide nucleic acid fluorescent in-situ hybridization (PNA FISH) can identify Candida species within 3 hours after a positive blood culture demonstrates yeast. A Candida albicans probe and a dual C. albicans/Candida glabrata probe are available in clinical practice, and multi-Candida species probes and flow cytometry are in development. Recent data suggest that the rapid identification of Candida species with PNA FISH can provide early targeted therapy to patients and thus reduce antifungal costs and improve patient care. This review evaluates the clinical and laboratory effects of the Candida probes and their limitations, and competing technologies. Candida PNA FISH probes offer clinicians the early knowledge of the species they are treating, allowing them to appropriately tailor therapy.

Rapid Molecular Diagnosis for Candida species Using PCR-RFLP

Rapid identification of Candida species in clinical laboratory is becoming increasingly important since the incidence of Candidiasis continues to rise as the hospital surveys show. Molecular techniques utilizing amplification of target DNA provide quick and precise methods for diagnosis and identification of Candida species. In this study, using universal primers, the ITS1-ITS4 region was amplified. The restriction enzyme MspI digests this region and was used to identify of C. albicans, C. glabrata, C. krusei, C. tropicalis and C. guilliermondii. Electrophoretically, ribosomal DNA of C. guilliermondii produced three bands whereas the other species gave two bands upon digestion. Accordingly these enzymes behave as valuable application tools in molecular diagnosis of Candida species in Candidiasis maladies and can be substituted with the classical diagnosis of the pathogen.

Rapid identification of Candida species in oral rinse solutions by PCR

Aims: To determine the sensitivity and specificity of a multiplex PCR assay for the contemporary identification of major species involved in oral candidiasis, without extraction and purification of DNA from...