Design of a Novel DNA-FISH Probe for the Rapid Identification of <i>Candida albicans</i> in Cosmetic Products

Microbiological quality control in cosmetic and pharmaceutical products is crucial for consumer safety. Traditional culture-based detection methods, such as plating on selective media, are time-consuming and may lack sensitivity. Fluorescence In Situ Hybridisation (FISH), a molecular and culture-ind...

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Bibliographic Details
Main Authors: Patrícia Branco, Margarida Nunes, Paula Pereira, Elisabete Muchagato Mauricio
Format: Article
Language:English
Published: MDPI AG 2025-06-01
Series:Cosmetics
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Online Access:https://www.mdpi.com/2079-9284/12/3/115
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Summary:Microbiological quality control in cosmetic and pharmaceutical products is crucial for consumer safety. Traditional culture-based detection methods, such as plating on selective media, are time-consuming and may lack sensitivity. Fluorescence In Situ Hybridisation (FISH), a molecular and culture-independent technique, enables rapid and precise microbial identification by targeting specific RNA or DNA sequences with fluorescent probes. In this study, a novel DNA-FISH probe was developed for the detection of <i>Candida albicans</i> in cosmetic formulations. The probe’s specificity was assessed in silico and experimentally using flow cytometry (flow-FISH) on <i>C. albicans</i> and non-target microorganisms, including <i>Pichia kudriavzevii</i>, commonly known as <i>Candida krusei</i>, <i>Saccharomyces cerevisiae</i>, <i>Wickerhamomyces anomalus</i>, <i>Escherichia coli</i>, and <i>Staphylococcus aureus</i>. The probe exhibited 98.9% hybridization efficiency with <i>C. albicans</i>, yielding a fluorescence intensity (FI) of 25,000 (a.u.), while non-target yeasts demonstrated minimal hybridization (4.7%, 2.3%, and 1.9% for <i>C. krusei</i>, <i>S. cerevisiae</i>, and <i>W. anomalus</i>, respectively) and bacteria showed negligible FI. Additionally, the probe’s applicability was evaluated in a tonic formulation, where <i>C. albicans</i>’ hybridization efficiency was slightly reduced to 88.4%, suggesting that formulation components may influence probe performance. Nevertheless, the probe maintained high specificity and efficiency without formamide, a toxic reagent commonly used in FISH. These findings highlight the potential of FISH probes for rapid, accurate, and safe microbial detection, offering a valuable tool for microbiological quality control in the cosmetics industry.
ISSN:2079-9284