Identification Of Candida Species Research Articles

Isolation, identification and antifungal susceptibility of Candida species in postmenopausal women

Abstract— The term "vulvovaginal candidiasis" refers to an inflammatory, non-erosive Vulvovaginitis characterized by symptoms such as itching, pain, dyspareunia, and discharge. The condition is treated with antifungal medication if a positive culture from a low vaginal swab reveals Candida. It should be mentioned that these people had ongoing medical conditions. VVC had previously been classified as acute or recurring. It appears that estrogen is a key permissive factor in VVC. Clinical findings have demonstrated that VVC is uncommon in premenopausal and postmenopausal women and that the post-menopausal vagina rarely contains isolated Candida. The study aimed to know the relationship between Vulvovaginitis and candida in postmenopausal women and effectiveness of some antifungals. A total of 100 postmenopausal women With Vulvovaginitis, aged 45 to 69, had vaginal swabs taken from them. Upon arrival at the laboratory, all samples were cultured on Sabouraud dextrose agar . The species of the isolated yeasts were identified by a variety of methods, including molecular analysis with primers ITS1–ITS4, growth on Chrom agar medium, and biochemical analyses. Result: candida in postmenopausal women is less than in per menopausal due to effectiveness of estrogen hormone in reproductive age . The outcomes On Chrom agar showed, six species of candida—C. albicans, C. glabrata, C. tropicalis, C. parapsilosis, C. krusei, and c. rugosa—were isolated. Among the other species, C. glabrata was discovered to be the most prevalent. The findings demonstrated that four species of yeast-like fungi were identified using molecular diagnosis and genetic testing as follows:

Phenotypic Identification And Genotypic Profiling Of Candida Species Isolated From Vaginal Swabs

Phenotypic Identification And Genotypic Profiling Of Candida Species Isolated From Vaginal Swabs

Highly Specific and Rapid Multiplex Identification of Candida Species Using Digital Microfluidics Integrated with a Semi-Nested Genoarray.

Candida species are the most common cause of fungal infections around the world, associated with superficial and even deep-seated infections. In clinical practice, there is great significance in identifying different Candida species because of their respective characteristics. However, current technologies have difficulty in onsite species identification due to long turnover time, high cost of reagents and instruments, or limited detection performance. We developed a semi-nested recombinase polymerase amplification (RPA) genoarray as well as an integrated system for highly specific identification of four Candida species with a simple design of primers, high detection sensitivity, fast turnover time, and good cost-effectiveness. The system constructed to perform the assay consists of a rapid sample processing module for nucleic acid release from fungal samples in 15 min and a digital microfluidic platform for precise and efficient detection reactions in 35 min. Therefore, our system could automatically identify specific Candida species, with a reagent consumption of only 2.5 μL of the RPA reaction mixture per target and no cross-reaction. Its detection sensitivity for four Candida species achieved 101-102 CFU/mL, which was 10-fold better than conventional RPA and even comparable to a common polymerase chain reaction. Evaluated by using cultured samples and 24 clinical samples, our system shows great applicability to onsite multiplex nucleic acid analysis.

Isolation and identification of Candida species from catheter-associated urinary tract infection in Thamar city hospitals, Yemen.

The incidence of urinary tract infections associated with Candida is increasing in Yemeni public hospitals. The primary objective of this research was to isolate specific Candida species responsible for catheter-associated urinary tract infections (UTIs) and to examine the antifungal sensitivity of these Candida isolates. A total of 200 samples were collected from patients with catheters admitted to multiple hospitals of Thamar city (Yemen). There were 50 positive samples with Candida out of 200 samples. We conducted the primary identification process using the established protocols. Before isolation and identification, all yeast isolates underwent sub-culturing on Sabouraud dextrose agar. We employed the standard microbiological procedures such as Gram staining, colonial morphology analysis, lactophenol cotton blue assay, germ tube formation assessment, colony staining on chrom agar Candida medium, and incubation at 37°C for 48h. The assessment of cultures was conducted by evaluating their predominant species. All Candida isolates were tested for antifungal susceptibility using the disk diffusion technique, as indicated by the Clinical and Laboratory Standards Institute (CLSI) M44-A document recommendations. In this study, the prevalence of Candida species obtained from catheter-associated UTIs was shown to be the highest among individuals aged 51-60years (28.0%) and the lowest was among those aged 10-20years (8.0%). Males exhibit higher rates than females, with males accounting for 56.0 and females for 44.0%, respectively. The predominant strain from catheters linked to urinary tract infections was Candida albicans. The Candida isolates had the highest susceptibility to itraconazole, with fluconazole and nystatin at sensitivity rates of 64, 60, and 50%, respectively. Amphotericin B and ketoconazole exhibited the most elevated concentrations. The p value of duration of catheterization < 5 was significant (p = 0.01), as well as significant in anti-fungal susceptibility testing of itraconazole, ketoconazole, and nystatin which are (p = 0.03), (p = 0.04), and (p = 0.03) respectively. Urinary tract infection due to candiduria was more common in patients with indwelling urinary catheter. The catheter-associated urinary tract infection caused by Candida species occurred mainly in old male patients. Candia albicans was the predominant Candida species isolated from urinary tract infection associated with urinary catheter at Thamar city hospitals. This study determined that diabetes and antibiotic use are significant predisposing factors associated with isolation of Candida in specimens submitted by patients at Thamar city hospitals.

Bridging Diagnostic Gaps: Utilising HiCrome Agar and Tetrazolium Reduction Medium for the Rapid and Presumptive Identification and Speciation of Candida Species in Vulvovaginal Candidiasis in Low-Resource Environments.

Background Vulvovaginal candidiasis (VVC) is a common fungal infection caused by an overgrowth of Candida species, primarily Candida albicans (C. albicans). Using HiCrome agar and tetrazolium reduction medium offers cost-effectiveness in Candida detection by eliminating the need for additional tests, reducing equipment costs compared to automated systems, and simplifying workflow with direct species identification while maintaining high specificity. They expedite detection by directly identifying Candida species based on colony colour, bypassing the multiple steps of phenotypic methods. This efficiency saves time in the laboratory, providing rapid results without the extended processing times associated with automated systems and facilitating prompt diagnosis and treatment decisions. These diagnostic tools are especially valuable in low-resource environments where a quick and accurate diagnosis of VVC is crucial for effective treatment and management of antifungal resistance. Aims and objectives This study aims to evaluate the efficacy of HiCrome agar and tetrazolium reduction medium's efficacy in speciating Candida species in VVC cases. Materials and methods A cross-sectional observational study was conducted at Saveetha Medical College and Hospitals, Chennai, India, over six months. High vaginal swabs from 126 patients suspected of VVC were collected and plated on Sabouraud dextrose agar (SDA), HiCrome Candida differential agar (Himedia, Mumbai, India), and tetrazolium reduction medium. The results were compared with those obtained from the VITEK2 compact system (bioMérieux, Marcy-l'Étoile, France). Results Of the 126 samples, 74.6% showed single yeast infections, 7.9% displayed mixed yeast infections, and 17.5% showed no growth. A total of 114 Candida isolates were identified. Both HiCrome agar and tetrazolium reduction medium accurately identified all isolates, with complete concordance with the VITEK2 compact system. The most commonly isolated species were C. albicans (55.2%), Candida tropicalis (32.4%), Candida glabrata (8.8%), and Candida parapsilosis (3.6%). Both media provided rapid and accurate presumptive identification in low-resource settings. Conclusions HiCrome agar and tetrazolium reduction medium demonstrated high sensitivity and specificity in identifying Candida species. These methods are reliable for rapid and accurate diagnosis, particularly in resource-limited settings. However, they may require supplementary tests for definitive species identification. The adoption of these diagnostic tools represents a significant advancement in clinical microbiology, improving VVC management and addressing antifungal resistance.

Vitek 2 system for identification and susceptibility testing of Candida species with detection of fluconazole resistant genes

Vitek 2 system for identification and susceptibility testing of Candida species with detection of fluconazole resistant genes

Oral Candida in post‐radiotherapy patients with xerostomia/hyposalivation: A narrative review

AbstractObjectiveHead and Neck Cancer (HNC) patients receiving radiotherapy (RT) often suffer from xerostomia and/or hyposalivation. As saliva plays an important antimicrobial and cleansing roles, these patients are at higher risks of opportunistic infections. This narrative review aims to provide an overview of current evidence on oral Candida colonisation and infection in these patients.MethodsA literature review of clinical studies on oral Candida colonisation and candidiasis in HNC patients receiving radiotherapy/chemoradiotherapy was conducted.ResultsMany clinical studies found high levels of Candida colonisation and a substantial proportion of post‐RT HNC patients suffering from oropharyngeal candidiasis (OPC). Importantly, oral Candida could be a reservoir for life‐threatening systemic infection in immunocompromised patients. The rising prevalence of non‐albicans Candida species and drug‐resistant infections has made identification of Candida species and antifungal susceptibility more important. Recent advances in oral microbiome and its interactions with Candida are discussed. This review also offers perspectives on limitations of current evidence and suggestions for future research.ConclusionFurther research to better understand Candida carriage, microbiome, OPC, and xerostomia/hyposalivation post‐RT would aid in devising a more comprehensive long‐term management plan and novel therapeutic approaches for HNC patients to achieve the full benefits of RT while minimising side effects.

Identification And Antifungal Susceptibility Testing Of Candida Species Isolated From Cancer Patients

Identification And Antifungal Susceptibility Testing Of Candida Species Isolated From Cancer Patients

Patients with recurrent vulvovaginal candidiasis exhibit a decrease in both the fungicidal activity of neutrophils and the proliferation of peripheral blood mononuclear cells.

Recurrent vulvovaginal candidiasis (RVVC) is an important and underestimated fungal infection. We aimed to determine the fungicidal and proliferative capacities of neutrophils and peripheral blood mononuclear cells (PBMCs), respectively and the clinical and microbiological characteristics of a cohort of Colombian patients diagnosed with RVVC. A cross-sectional study was conducted. A total of 66 women were included (40 diagnosed with RVVC and 26 healthy women [HW]). Demographic and clinical data were recorded. Vaginal fluid samples were obtained for isolation, identification and antifungal susceptibility testing of Candida species using selective culture media and the Vitek 2.0® system. Blood samples were also obtained to evaluate cell subpopulations; furthermore, neutrophils and PBMCs were isolated to determine their fungicidal and proliferative capacities, respectively. The median age was 29 (IQR: 34-23) for RVVC and 24 (IQR: 30-23) for HW. Only two species of the genus Candida were identified: Candida albicans (92.5%) and Candida lusitaniae (7.5%). Resistance to fluconazole, voriconazole, flucytosine and amphotericin B was observed on six C. albicans isolates and one C. lusitaniae isolate. Only the family history of vulvovaginal candidiasis was associated with RVVC occurrence. The RVVC group exhibited a significantly higher number of neutrophils but with lower fungicidal activity in comparison to HW; likewise, PBMCs from RVVC patients presented a lower proliferation index when stimulated with C. albicans. Contrary to what has been reported worldwide, in Colombian patients with RVVC, C. albicans was the main isolated species without increased antifungal resistance. The diminished fungicidal and proliferative capacities of neutrophils and PBMCs, respectively, could suggest a possible alteration in the innate and adaptive immune responses.

Rapid identification,fluconazole and micafungin susceptibility testing of Candida species from blood culture by a short incubation method

This study aimed to develop and validate a rapid method for identification by MALDI-TOF system and determination of the susceptibility to Fluconazole and Micafungin by broth microdilution among Candidaspecies causing bloodstream infections. Subcultures from blood culture bottles were incubated for 5 hours (+/- 1h) and used to perform the tests, so that the turnaround time of rapid identification and susceptibility profile was about 5 and 24 hours, respectively. The rapid identification showed agreement of 92.05 %. Regarding the rapid broth microdilution for Fluconazole and Micafungin, the agreement was 97.06 % (p<0.001) and 100 % (p<0.001), and the Kappa coefficient was 0.91 (p<0.001) and 1.0 (p<0.001), respectively. To conclude, both rapid methods showed to be reproducible, inexpensive, easy to perform and time-saving. Thus, these methodologies could be useful to guide and adjust empirical antifungal therapy.

Prevalence of Candida Species Among Unmarried Women Attending Lafia Primary Health Care Centers

One of the several normal-flora of an individual’s vaginal, oral cavity, gastrointestinal tract and mucosal membrane are Candida species. Candida have been associated with diverse diseases including life threatening invasive, superficial and hematogenic infections. This study determined the prevalence of Candida species amongst unmarried women attending Lafia Primary Health Care Centers. Forty high vagina swabs were collected from unmarried women. Isolation and identification of Candida species were performed according to standard procedures using Sabouraud Dextrose Agar, Gram staining and Germ tube test. 13 (32.5%) were positive for five different Candida species. C. albicans was the most prevalent (38.46%; 5/13) species while non-albicans Candida [C. glabrata (23.07%; 3/13), C. tropicalis (6.89%; 2/13), C. parapsilosis (3.44%; 1/13) and C. krusei (6.89%; 2/13)] accounted for 61.54% (8/13) of the total positive cases. Women aged 15-18 years and 23-26 years had the lowest and highest prevalence rates of 14.28% (1/7) and 42.10% (8/19) respectively. Candidiasis is a threat to global health and should be taken with all seriousness. The high candidiasis prevalence observed herein portrays a concern and calls for a continuous study of unmarried women attending Primary Healthcare Centers in Lafia. Furthermore, there should be an improvement in creating awareness of the opportunistic pathogenic characteristic of Candida species as normal flora and the effect they cause when there are hormonal imbalances caused by drug abuse, pregnancy, and poor dieting. The practice of good hygiene should be observed to prevent the spread of the infection.

Speciation of Candida species isolated from cutaneous skin lesions using CHROMagar and conventional methods

Background: Candida species are members of the normal flora of the skin, mucous membranes, and gastrointestinal tract. They are endogenous opportunists which cause secondary infection in individuals with some underlying immunocompromised conditions. Candidiasis is a common fungal disease found in humans affecting mucosa, skin, nails, and internal organs of the body. Candida albicans is generally considered the major pathogen among the Candida species. An increase in the prevalence of non-albicans species has been noted during the last decades. Aims and Objectives:The study was done to evaluate the performance of the conventional identification method and commercially available chromogenic Candida speciation media (CHROMagar) for the identification of medically important yeast and yeast-like organisms in a routine clinical microbiology laboratory. Materials and Methods: This study was conducted in the Department of Microbiology, Gandhi Medical College, Hamidia, and Associated Hospitals, Bhopal (M.P.), from January 2020 to October 2021. A total of 180 samples were collected from patients suspected of superficial mycoses which were then subjected to direct KOH examination and culture on plain Sabouraud’s dextrose agar (SDA), on SDA with antibiotics slants (cycloheximide and chloramphenicol). The growths were examined macroscopically as well as microscopically. Germ tube tests and inoculations on CHROMagar Candida medium were performed on all growths identified as yeasts and confirmed by morphology on CMA. Results: In our study, the majority of patients, belonged to the age group of 21–30 years (25%) with a male-to-female ratio of 2:1. Candida tropicalis (44.68%) was the most common Candida species, followed by C. albicans (38.30%), Candida krusei (12.77%), and Candida glabrata (4.25%). Conclusion: Along with C. albicans, non-albicans Candida spp such as C. tropicalis, C. krusei, and C. glabrata are increasingly being isolated from clinical specimens. Identification of Candida species by colony color on Hicrome correlated with the morphology on CMA. Thus, the incorporation of CMA in routine yeast identification is more judicious than Hicrome as it increases the accuracy in the identification of Candida species within the same time as that of Hicrome.

Molecular Detection and Identification of Candida Species Isolates from Oral by RFLP-PCR

In this study, 10 local isolates from a total of 50 samples of Candida sp. were collected from oral swabs of patients with oral infections in Mosul hospitals. The isolates were diagnosed based on culturing, microscopic and biochemical characteristics, and then molecular methods. The first diagnosis by culturing, microscopic and biochemical tests found the isolates were identified as Candida sp. The ITS region was amplified using universal primers (ITS4-ITS5), The PCR product was size (510-721) bp. Performing RFLP-PCR using MspI, HhaI,and EcoRI, restriction enzyme to detect and identify Candida species, the results showed the presence of the cutting sequence of MspI and HhaI enzymes in the genomic DNA content of local isolate and the absence of the sequences for the EcoRI restriction enzyme. Two Candida species were identified (C. krusei and C. the basis of size and fragment sequences then compared with sequences of standard strains from the gene bank in previous studies. Therefore, it can be observed that there is a genetic variation between the local isolates and that there are different genotypes of rDNA 5.8S have been diagnosed in 10 isolates after the cutting process with three restriction enzymes. We conclude from this study that the RFLP-PCR technique was the best in diagnosing and identifying Candida species compared with traditional methods. and we are d the genetic variation between local isolates.

Performance of Chromogenic Candida Lab-Agar® Medium in Presumptive Identification of Candida Species from Clinical Samples at Sourô Sanou University Hospital of Bobo-Dioulasso, Burkina Faso

Introduction: The incidence of Candida infections is increasing worldwide. In clinical laboratories of resource-constrained countries, Candida speciation is commonly limited to germ tube tests and culture onto a chromogenic medium. In this study, we evaluated the diagnostic performance of Chromogenic Candida Lab-Agar® (CCL) in identifying Candida species from clinical samples. Methods: We evaluated the diagnostic performance of CCL with 83 yeast isolates collected from 73 clinical samples at the laboratory department of Sourô Sanou University Hospital of Bobo-Dioulasso, Burkina Faso. Clinical specimens included vaginal swabs, urine, and blood cultures. After preliminary isolation on Sabouraud chloramphenicol agar, yeast isolates were inoculated onto the CCL medium and incubated at 35 °C for 48 h. Matrix-assisted laser desorption/ionisation time of flight mass spectrometry (MALDI-TOF MS) and ribosomal DNA internal transcribed spacer (ITS) sequencing were used as reference methods. Results: Among yeast species, Candida albicans was the most prevalent (43.4%), followed by C. krusei (13.3%), C. glabrata (12.0%), C. kefyr (8.4%), and C. tropicalis (7.2%). The overall agreement rate of CCL was 56.6% and varied across Candida species; it was 94.4% for C. albicans, 50% for C. glabrata, 18.2% for C. krusei, and 33.3% for C. tropicalis. Conclusions: This study showed that CCL had moderate accuracy in identifying Candida at the species level from clinical specimens in a routine laboratory in Burkina Faso. The misidentification of non-albicans species may expose patients to inadequate antifungal treatment. Therefore, identifying yeast in a routine based on CCL is not enough and should be associated with more accurate methods.

Exploring Molecular Methods for Candida species Identification and Characterization

Candida species (CS) are significant fungal pathogens responsible for a wide range of infections in humans, making their precise and timely identification crucial for effective treatment and disease management. Traditional identification methods, such as phenotypic assays and culture-based techniques, often suffer from limitations, including prolonged processing times, limited accuracy, and inadequate differentiation between species. As a result, molecular techniques have gained prominence for their ability to rapidly and accurately identify and characterize various Candida species. This review discusses the shortcomings of conventional methods and highlights the array of molecular strategies that have been developed, such as polymerase chain reaction (PCR) assays, DNA sequencing, and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS). These techniques leverage the unique genetic and proteomic profiles of Candida species to provide highly sensitive and specific identification, even distinguishing between closely related strains. Additionally, the review explores the use of molecular approaches in clinical diagnostics, epidemiological studies, and antifungal resistance monitoring, where they enable the rapid detection of Candida species from clinical samples, allowing for swift diagnosis and timely antifungal intervention. Furthermore, these techniques are instrumental in identifying clonal outbreaks and tracking transmission dynamics in healthcare settings. The review emphasizes the advantages of molecular methods—such as their precision, speed, and capacity to uncover cryptic species—while also calling for continued research to improve their efficacy and broader application in Candida species identification.

Candidaemia: A 9-Year Retrospective Analysis of Epidemiology and Antimicrobial Susceptibility in Tertiary Care Hospitals in Western China.

This investigation endeavors to scrutinize the resistance profiles to antifungal agents, alongside the clinical distribution of Candida isolates that yielded positive results in blood cultures at Suining Central Hospital spanning the years 2015 to 2023. The objective is to provide crucial epidemiological insights that may aid in early clinical intervention and judicious deployment of antifungal therapies. This retrospective analysis analyses data on 182 different Candida strains with positive clinical blood cultures obtained from the Microbiology Laboratory of Suining Central Hospital over a period of nine consecutive years. The study involved identification of Candida species and assessment of resistance patterns to fungal drugs. Our analysis revealed that the median age of patients diagnosed with Candidaemia from the 182 strains was 62 years, with a distribution of 63.7% females and 36.3% males. Within the cohort of 182 Candida strains, Candida albicans constituted 32.4%, while non-albicans Candida species comprised 67.6% of the cases. Specifically, Candida tropicalis represented 37.4%, Candida glabrata 12.1%, Candida parapsilosis 11.0%,Candida guilliermondii 3.8%, and both Candida krusei and Candida Dublin accounted for 1.6% each. These Candida species were predominantly identified in intensive care units (ICU), hematology, gastroenterology, neurology centers, and endocrine metabolism units. The findings of this investigation suggest a shift in the prevalence of non-Candida albicans species, notably C. tropicalis, as the predominant cause of Candidaemia at Suining Central Hospital, surpassing C. albicans. Although instances of antifungal resistance are infrequent, there has been a notable rise in resistance to azoles. This study provides important insights into the local epidemiology, which will be essential for informing the selection of empirical antifungal therapy and contributing to the global surveillance of antifungal resistance.

Candida species isolated in female patients of reproductive age with vaginal candidiasis in Gualeguaychú, Entre Ríos, Argentina

Objective: To identify the Candida species isolated in female patients of reproductive age with vaginal candidiasis. To determine the antifungal (ATF) sensitivity profile of the isolated Candida species. Materials and methods: Cross-sectional descriptive study, in which 124 Candida spp strains from vaginal discharge samples were isolated in Sabouraud medium supplemented with glucose. CHRO Magar Candida was used for species identification and complementary tests such as germ tube, chlamydoconidia development and investigation of trehalose assimilation were performed. In vitro sensitivity was investigated by diffusion method with ATF discs fluconazole (FLU), itraconazole (ITRA) and amphotericin B (AMB). Results: C. albicans was observed in 85.5% of the isolates followed by C. glabrata with 7.3%, C. krusei 4%, C. tropicalis 2.4% and other Candida species. For FLU, 0.9% of C. albicans, 11.1% of C. glabrata and 100% of C. krusei isolates showed resistance. For ITRA, 17% of C. albicans isolates, 55.6% of C. glabrata and 100% of C. krusei were resistant. There were no isolates resistant to AMB. Conclusion: Vaginal candidiasis continues to present Candida albicans as the main etiological agent, which is widely sensitive to ATFs. C. glabrata and C. krusei species show increased resistance to azoles. The results obtained ratify the growing need for Candida species identification tests and determination of in vitro sensitivity to ATFs in order to guide the treatment of vaginal candidiasis towards therapeutic success.

Phenotypic characterization and antifungal susceptibility profile of Candida isolates from women with vulvovaginal candidiasis at a community health center linked to a teaching institution in the Sub-Himalayan region of North India.

Vulvovaginal candidiasis (VVC) is considered a common gynecological problem among females of reproductive age group. 70-75% of women report having had candidal vulvovaginitis at some point in their lifetimes and 40-50% suffer recurrent candidal vulvovaginitis. This study aims to identify the Candida species involved in VVC and to determine their antifungal susceptibility pattern. The present study was a cross-sectional study conducted on 257 females (18-55 yr) with complaints of abnormal vaginal discharge. For detection of Candida, the swab samples were subjected to Gram stain, 10% KOH mount, and culture on Sabouraud dextrose agar (SDA). Candida species identification was done by subculturing Candida isolates onto CHROMagar, corn meal agar (Himedia), and further confirmation was done by MALDI-TOF MS. Antifungal testing was done using the disk diffusion method. A total of 257 females with complaints of abnormal discharge were enrolled in this study. Out of 257, C. albicans 37 (58.7%) and 26 (41.3%) isolates were identified as non-albicans Candida. Out of 63 positive cases, a maximum number of study subject belongs to the age group 26-35 years (50.8%). Along with vaginal discharge, itching (65.37%) is the most common complaint. VVC was found to be most predominant in patients with prolonged antibiotic therapy (38.1%), and in pregnant females (15.9%). Understanding the emerging fungal pathogens and their drug susceptibility patterns is essential for the effective management of infections. Drug resistance can lead to treatment failure and highlights the need for alternative treatment options or strategies.

620. Development and Validation of a Novel qPCR Assay for Differentiation of Common Candida Pathogens including C. auris

Abstract Background Candida is a common asymptomatic fungal colonizer of humans. Of the 200+ total species in the genus Candida, only a few are known to cause infection. The most common are C. albicans, C. glabrata, C. parapsilosis, C. tropicalis, C. krusei, and C. auris. In the USA, candidemia is the fourth most common cause of bloodstream infections in hospitalized patients. Although antifungal medication can treat candidemia, mortality rates remain between 30 and 60%. Eurofins Viracor has developed a qPCR assay that will qualitatively detect and differentiate any of the 6 previously listed Candida species in whole blood. Speciation is critical for treatment due to variable drug susceptibility among different species, particularly C. auris due to its multi-drug resistance. Methods Multiple primer/probe mixes were designed around and within the ITS1, 5.8s, ITS2, and 28s ribosomal subunit regions in each of the six Candida target species. The 5.8s and 28s regions are heavily conserved, while the ITS regions are highly divergent and used to differentiate species. After optimization, the six primer/probe mixes were partially multiplexed resulting in four mixes covering all species. DNA was extracted from human blood spiked with viable Candida using the MagMAX™ DNA Multi-Sample Ultra 2.0 Kit, and eluates were amplified using the Applied Biosystems™ 7500 Fast platform. Results Each assay met all validation acceptance criteria and demonstrated excellent accuracy, precision, linearity and dynamic range, and sensitivity. 32 non-Candida fungal and bacterial pathogens that commonly cause bloodstream infections were used to assess specificity. All specificity samples were negative for all Candida targets. To assess accuracy, 75 of 75 Candida samples representing all six species across a range of concentrations were tested and correctly identified. 24 of 25 known-negative samples were negative, with one positive C. glabrata CT result observed just below the cutoff. Conclusion Traditional blood culture methods for Candida species identification can be too time consuming for a quickly progressing infection. The Candida qPCR assay described here not only detects an infection faster but provides accurate speciation allowing for rapid and targeted antifungal treatment options for the patient. Disclosures Clayton Thomas, BS in Biology, Eurofins Viracor: Employee James Grantham, n/a, Eurofins Viracor: Employee Emily Lute, n/a, Eurofins Viracor: Employee Jerod Davidson, n/a, Eurofins Viracor: Employee Scott Cowden, MS, MB ASCP(CM), Eurofins Viracor: Employee Jamie Nutt, n/a, Eurofins Viracor: Employee

Phenotypic identification of Candida species and relative expression of candida drug resistance (CDR1) gene in fluconazole resistant and sensitive Candida albicans

ABSTRACT:&#x0D; Background and objective:&#x0D; Candida albicans belongs to the genus Candida. It is a commensal yeast in healthy individuals and can cause wide variety of infections in immune-compromised patients. The aim of the present study was to observe the frequency of Candida species, resistant profile of C. albicans against fluconazole and over-expression of CDR1 gene in both fluconazole resistant and sensitive C. albicans.&#x0D; Methodology:&#x0D; A total number of 206 Candida species were collected on Sabouraud Dextrose agar from Jinnah hospital Lahore. The isolates were re-confirmed by using various microbiological techniques like wet preparation, Gram stain and Germ tube test. Identification upto the species level was done by APIC 20AUX. Antifungal susceptibility profile of C. albicans against fluconazole was determined by broth microdilutiom method. Real Time PCR was performed on both the sensitive and resistant C. albicans to observe the role of CDR1 gene against fluconazole resistance. Results:&#x0D; &#x0D; tropicals was the most common species with the frequency of 66 (32.0%) followed by C. albicans 41(19.9), C. glabrata 24 (11.7%), C. lusitaniae 14 (6.8%) and other less common species. The majority of infected patients were females (59.2%) than males (41%). Antifungal susceptibility testing showed that 80% C. albicans were sensitive to fluconazole. Results of gene expression showed that CDR1 gene play a major role towards fluconazole resistance in C.albicans.Conclusion: Fluconazole was found to be effective in treating infections caused by C. albicans. &#x0D; &#x0D; Keywords: Candida albicans, Candida Drug Resistant Gene 1, Polymerase Chain Reaction, Candida Tropicalis, Candida glabrata, and Candida lusitaniae