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Molecular Identification of Candida Species among Iranian Patients: Pursuing Candida auris
Fatemeh Safari1, Mahboobeh Madani1, Farshad Zare2
1Department of Microbiology, Fal.C., Islamic Azad University, Isfahan, Iran.
Background:
Candida species are opportunistic pathogens that can cause local or systemic invasive infections in patients hospitalized in intensive care units (ICUs). The present study aimed to rapidly and precisely identify Candida species, especially Candida auris, from patients hospitalized in high-risk units using the polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) technique.
Materials And Methods:
In this study, 136 clinical samples were prospectively collected from different organs of patients hospitalized in high-risk units recruited from three tertiary care training hospitals in Isfahan. Direct microscopic analysis using potassium hydroxide 10% followed by culturing on Sabouraud dextrose agar and CHROMagar culture media was primarily used to screen Candida species. Molecular detection of clinical isolates was mainly done using the PCR-RFLP method. Then, all the isolates were subjected to confirmatory diagnosis again through amplification of the C. auris-specific PCR.
Results:
Candida albicans was the most prevalent isolate (48.8%), followed by Candida glabrata (17.6%), Candida tropicalis (16.1%), Candida parapsilosis (6.6%), Candida krusei (5.8%), Candida famata (4.4%), Candida kefyr (2.9%), and Candida guilliermondii (1.4%). Therefore, the prevalence of non-albicans Candida species was higher than C. albicans isolates. No C. auris isolates were identified among 136 patients. However, according to the positive control, C. auris had no restriction enzyme cleavage site for MspI, and its PCR and digestion products were the same size (401 bp).
Conclusions:
The simple and reliable PCR-RFLP assay used in the study has the potential to identify rare yeasts such as C. auris. It can reduce turnaround times and costs if applied in developing countries.
Insights
This study evaluated the PCR-restriction fragment length polymorphism (PCR-RFLP) technique for identifying Candida species, including Candida auris, in high-risk patients. While Candida albicans was most common, no C. auris was detected, but the method showed potential for rapid yeast identification.
Area of Science:
- Medical Mycology
- Clinical Microbiology
- Molecular Diagnostics
Background:
- Candida species are opportunistic pathogens causing infections in intensive care unit (ICU) patients.
- Accurate and rapid identification of Candida species, particularly Candida auris, is crucial for effective patient management.
- High-risk units require timely diagnostic methods to combat invasive fungal infections.
Purpose of the Study:
- To assess the efficacy of the polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) technique for identifying Candida species.
- To specifically target the rapid and precise detection of Candida auris in clinical samples from high-risk patients.
- To evaluate the prevalence of different Candida species in hospitalized patients within high-risk units.
Main Methods:
- Prospective collection of 136 clinical samples from patients in high-risk units across three tertiary care hospitals.
- Initial screening using direct microscopy (10% KOH) and culture on Sabouraud dextrose agar and CHROMagar.
- Molecular identification primarily employed PCR-RFLP, with confirmatory diagnosis using C. auris-specific PCR.
Main Results:
- Candida albicans was the most frequent isolate (48.8%), followed by non-albicans species like Candida glabrata (17.6%) and Candida tropicalis (16.1%).
- Non-albicans Candida species collectively showed higher prevalence than Candida albicans.
- No Candida auris isolates were identified in the 136 patient samples; however, the PCR-RFLP method confirmed its characteristic banding pattern.
Conclusions:
- The PCR-RFLP assay is a simple, reliable method for identifying Candida species, including rare yeasts like C. auris.
- This molecular technique has the potential to reduce diagnostic turnaround times and costs.
- The application of PCR-RFLP in developing countries could significantly improve the management of invasive fungal infections.
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