Quantification of Malassezia pachydermatis by real-time PCR in swabs from the external ear canal of dogs
Puig Sanz, Lluís. (Universitat Autònoma de Barcelona. Departament de Sanitat i d'Anatomia Animals)
Castellá, Gemma (Universitat Autònoma de Barcelona. Departament de Sanitat i d'Anatomia Animals)
Cabañes Sáenz, Francisco Javier (Universitat Autònoma de Barcelona. Departament de Sanitat i d'Anatomia Animals)

Data:	2019
Resum:	Malassezia pachydermatis is part of the normal microbiota of canine skin and external ear canal, and is also associated with otitis externa in dogs. Laboratory detection of Malassezia otitis relies on the presence of elevated numbers of the yeast on cytologic examination of otic exudate. Although cytology has high specificity, it has low sensitivity, resulting in false-negatives and posing a challenge for clinicians to accurately diagnose Malassezia otitis. We developed a quantitative PCR (qPCR) to detect and quantify M. pachydermatis yeasts and validate the method with swabs from external ear canals of dogs. Our qPCR uses the β-tubulin gene, a single-copy gene, as a target. The limit of quantification was established as 0. 18 ng/reaction, equivalent to 2. 0 × 10 genome equivalents (gEq). Swabs from healthy dogs yielded quantification values of ≤2. 7 × 10 gEq in the qPCR, whereas swabs from dogs with otitis yielded quantification values of ≥2. 5 × 10 gEq. Our qPCR assay provides accurate quantification of M. pachydermatis yeasts from swab samples from dogs, is more sensitive than cytology, and could be used to monitor response to treatment. Our assay could also be valuable in a research setting to better understand the pathogenesis of M. pachydermatis.
Nota:	Financial support for this project came from Servei Veterinari de Bacteriologia i Micologia from the Universitat Autònoma de Barcelona.
Drets:	Aquest document està subjecte a una llicència d'ús Creative Commons. Es permet la reproducció total o parcial, la distribució, la comunicació pública de l'obra i la creació d'obres derivades, fins i tot amb finalitats comercials, sempre i quan es reconegui l'autoria de l'obra original.
Llengua:	Anglès
Document:	Article ; recerca ; Versió publicada
Matèria:	Gossos ; Otorinolaringoloria veterinària ; Quantitative PCR ; Otitis ; β-tubulin ; Malassezia pachydermatis
Publicat a:	Journal of Veterinary Diagnostic Investigation, Vol. 31 Núm. 3 (january 2019) , p. 440-447, ISSN 1943-4936

DOI: 10.1177/1040638719840686
PMID: 30943876

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