Evaluation of an in-house pan-Malassezia quantitative PCR in human clinical samples

Euzen, Victor Data curation; Formal analysis; Validation; Visualization; Writing - original draft; Writing - review & editing
Ghelfenstein-Ferreira, Théo Conceptualization; Data curation; Formal analysis; Investigation; Methodology; Resources; Validation; Visualization; Writing - review & editing
Benhadid-Brahmi, Yasmine Data curation; Writing - review & editing
Teboul, Alexandra Data curation; Writing - review & editing
Dellière, Sarah Conceptualization; Writing - review & editing
Benderdouche, Mazouz Data curation; Writing - review & editing
Charlier, Véronique Data curation
Desnos-Ollivier, Marie Data curation; Writing - review & editing
Hamane, Samia Conceptualization; Methodology; Resources; Validation; Writing - review & editing
Alanio, Alexandre Conceptualization; Funding acquisition; Methodology; Project administration; Supervision; Validation; Writing - review & editing

Mycology and Parasitology Department, Hôpital Saint-Louis, Assistance Publique-Hôpitaux de Paris, 1 av. Claude Vellefaux, 75010 Paris, France
Mycology and Parasitology Department, Hôpital Saint-Louis, Assistance Publique-Hôpitaux de Paris, 1 av. Claude Vellefaux, 75010 Paris, France
Mycology and Parasitology Department, Hôpital Saint-Louis, Assistance Publique-Hôpitaux de Paris, 1 av. Claude Vellefaux, 75010 Paris, France
Mycology and Parasitology Department, Hôpital Saint-Louis, Assistance Publique-Hôpitaux de Paris, 1 av. Claude Vellefaux, 75010 Paris, France
Mycology and Parasitology Department, Hôpital Saint-Louis, Assistance Publique-Hôpitaux de Paris, 1 av. Claude Vellefaux, 75010 Paris, France
Mycology and Parasitology Department, Hôpital Saint-Louis, Assistance Publique-Hôpitaux de Paris, 1 av. Claude Vellefaux, 75010 Paris, France
Mycology and Parasitology Department, Hôpital Saint-Louis, Assistance Publique-Hôpitaux de Paris, 1 av. Claude Vellefaux, 75010 Paris, France
Natl. Ref. Center for Invasive Mycoses and Antifungals, Institut Pasteur, 25 Rue du Dr Roux, 75015 Paris, France
Mycology and Parasitology Department, Hôpital Saint-Louis, Assistance Publique-Hôpitaux de Paris, 1 av. Claude Vellefaux, 75010 Paris, France
Mycology and Parasitology Department, Hôpital Saint-Louis, Assistance Publique-Hôpitaux de Paris, 1 av. Claude Vellefaux, 75010 Paris, France
Natl. Ref. Center for Invasive Mycoses and Antifungals, Institut Pasteur, 25 Rue du Dr Roux, 75015 Paris, France

To whom correspondence should be addressed: Alexandre Alanio, MD, PhD, Tel: +33140613255; E-mail: [email protected]

Received January 17, 2024

Received in revised form August 16, 2024

Accepted September 11, 2024

Revised October 04, 2024

This article is published and distributed under the terms of the Oxford University Press, Standard Journals Publication Model (https://academic.oup.com/journals/pages/open_access/funder_policies/chorus/standard_publication_model)

Medical Mycology 62(10), October 2024. | DOI: 10.1093/mmy/myae095

Lay Summary

Malassezia species are ubiquitous members of various human microbiomes, including cutaneous and mucosal sites. While these fungi have been implicated in several pathologies, their presence as commensals complicates their study, especially due to difficulties in culturing them in vitro. This has necessitated the development of molecular techniques to detect and quantify Malassezia species directly from clinical samples.

In this study, we report on the development and application of an in-house pan-Malassezia quantitative PCR (panM-qPCR) assay. This assay targets the conserved 28S rDNA gene across all known Malassezia species, allowing for a broad-spectrum detection approach. We applied this panM-qPCR to a diverse set of clinical samples, totaling 361 specimens from 161 subjects, encompassing healthy individuals, patients with seborrheic dermatitis, burn victims, and immunocompromised individuals.

Our results indicate variable Malassezia loads on different skin sites of healthy volunteers, with significantly lower quantification cycle (Cq) values observed in seborrheic dermatitis patients, suggesting an increased fungal burden. Burn patients also showed a marked increase in Malassezia spp. levels compared to healthy individuals. Stool samples demonstrated a low prevalence (6.7%) of Malassezia spp., but with high Cq values when present. Notably, ear samples revealed a higher positivity rate compared to nasal samples.

The findings highlight the practicality and sensitivity of qPCR for elucidating the Malassezia burden across various human samples. This molecular approach confirms the differential colonization of Malassezia spp. in different clinical contexts. The panM-qPCR offers a promising approach for comprehensive mycobiota research, particularly in conditions where culture-based methods fall short.