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A 96 Well Microtiter Plate-based Method for Monitoring Formation and Antifungal Susceptibility Testing of Candida albicans Biofilms
Published on: October 21, 2010
Robust Fluorometric Aptamer Assay for Direct and Rapid Detection of Clinical Isolates of Candida spec
Yiting Zhang1, Hu Xing1, Grigory Bolotnikov1
1Institute of Pharmaceutical Biotechnology, Ulm University, Albert-Einstein-Allee 11, 89081 Ulm, Germany.
Abstract:
Infections caused by yeasts of the genus Candida are likely to occur not only in immunocompromised patients but also in healthy individuals, leading to infections of the gastrointestinal tract, urinary tract, and respiratory tract. Due to the rapid increase in the frequency of reported Candidiasis cases in recent years, diagnostic research has become the subject of many studies, and therefore, we developed a polyclonal aptamer library-based fluorometric assay with high specificity and affinity towards Candida spec. to quantify the pathogens in clinical samples with high sensitivity. We recently obtained the specific aptamer library R10, which explicitly recognized Candida and evolved it by mimicking an early skin infection model caused by Candida using the FluCell-SELEX system. In the follow-up study presented here, we demonstrate that the aptamer library R10-based bioassay specifically recognizes invasive clinical Candida isolates, including not only C. albicans but also strains like C. tropcialis, C. krusei, or C. glabrata. The next-generation fluorometric bioassay presented here can reliably and easily detect an early Candida infection and could be used for further clinical research or could even be developed into a full in vitro diagnostic tool.
Insights
A new aptamer-based assay accurately detects Candida infections in clinical samples. This sensitive fluorometric bioassay identifies various Candida species, aiding early diagnosis.
Area of Science:
- Clinical microbiology
- Biotechnology
- Infectious diseases
Background:
- Candida yeast infections affect both immunocompromised and healthy individuals.
- Rising Candidiasis cases necessitate improved diagnostic methods.
- Current diagnostics may lack the sensitivity for early detection.
Purpose of the Study:
- To develop a highly specific and sensitive assay for Candida detection.
- To create a fluorometric bioassay using an aptamer library.
- To validate the assay's performance on clinical isolates.
Main Methods:
- Development of a polyclonal aptamer library (R10) targeting Candida.
- Evolution of the aptamer library using the FluCell-SELEX system.
- Creation of a fluorometric bioassay for pathogen quantification.
Main Results:
- The R10 aptamer library demonstrated high specificity and affinity for Candida.
- The aptamer-based bioassay successfully recognized various clinical Candida isolates, including C. albicans, C. tropicalis, C. krusei, and C. glabrata.
- The assay showed potential for reliable and easy early Candida infection detection.
Conclusions:
- A novel aptamer library-based fluorometric assay offers a sensitive method for detecting Candida.
- This next-generation bioassay can identify diverse clinical Candida species.
- The developed tool holds promise for clinical research and potential in vitro diagnostic applications.
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