Multiple region high resolution melting-based method for accurate differentiation of food-derived yeasts at species level resolution

cris.virtual.author-orcid0000-0002-8601-3536
cris.virtual.author-orcid0000-0001-8372-8459
cris.virtualsource.author-orcidd5dfe19f-4d96-409d-9404-27a4f41fdb18
cris.virtualsource.author-orcid6f5a4155-2edb-48cf-b362-f3180e151169
dc.abstract.enYeasts comprise a divergent group of microorganisms playing an important role in foods production. To monitor the production processes, validate authenticity and safety of foods, reliable methods of yeast identification and differentiation are needed. Nowadays, traditional PCR/sequencing-based methods are replaced by rapid tech niques not requiring post-PCR processing. In the present study, we developed a three region-based qPCR-HRM protocol, for rapid differentiation of yeast species isolated from food products. The three targeted fragments (26S rDNA, 18S rDNA, ITS) were carefully analyzed to dock the primers at inter-species conservative regions, flanking polymorphic regions of ~200 bp. Thirty eight yeast strains were used as a training material. The collection of yeast spanned Pichia, Clavispora, Candida, Yarrowia, Kluyveromyces, Saccharomyces, and Wickerhamomyces genera. MALDI-TOF mass spectrometry and conventional rDNA sequencing were used for validation. Conducted studies demonstrated that when used individually, each of the three regions analyzed by qPCR-HRM possessed its own yeast species-specific limitations, sometimes leading to inaccurate taxonomic classification. On the other hand, simultaneous analysis of the three proposed regions resulted in rapid and adequate differentiation of all the yeast strains at species-level resolution.
dc.affiliationWydział Nauk o Żywności i Żywieniu
dc.affiliation.instituteKatedra Biotechnologii i Mikrobiologii Żywności
dc.contributor.authorBorkowska, Monika
dc.contributor.authorCelińska, Ewelina
dc.date.access2025-06-18
dc.date.accessioned2025-09-30T10:55:22Z
dc.date.available2025-09-30T10:55:22Z
dc.date.copyright2022-08-26
dc.date.issued2023
dc.description.accesstimeat_publication
dc.description.bibliographyil., bibliogr.
dc.description.financepublication_nocost
dc.description.financecost0,00
dc.description.if4,5
dc.description.numberFebruary 2023
dc.description.points140
dc.description.versionfinal_published
dc.description.volume109
dc.identifier.doi10.1016/j.fm.2022.104120
dc.identifier.eissn1095-9998
dc.identifier.issn0740-0020
dc.identifier.urihttps://sciencerep.up.poznan.pl/handle/item/5045
dc.identifier.weblinkhttp://www.sciencedirect.com/science/article/pii/S0740002022001447
dc.languageen
dc.relation.ispartofFood Microbiology
dc.relation.pagesart. 104120
dc.rightsCC-BY-NC-ND
dc.sciencecloudnosend
dc.share.typeOTHER
dc.subject.enfungi
dc.subject.enidentification
dc.subject.enrDNA sequencing
dc.subject.enQPCR
dc.subject.enHRM analysis
dc.subject.enMALDI-TOF mass spectrometry
dc.titleMultiple region high resolution melting-based method for accurate differentiation of food-derived yeasts at species level resolution
dc.typeJournalArticle
dspace.entity.typePublication
oaire.citation.volume109