title: Development and application of a quantitative real-time PCR assay for rapid detection of the multifaceted yeast Kazachstania servazzii in food.
abstract: The beneficial contributions of Kazachstania servazzii are well-established in various food processes. This yeast also contributes in the spoilage of finished packaged food due to abundant gas production. In particular, an occurrence of K. servazzii was recently positively correlated with the formation of severe package swelling of some prepared fresh pizzas. To circumscribe this concern, a quantitative SYBR green real-time PCR assay based on a newly designed specific primer pair targeting the ribosomal ITS1-5.8S-ITS2 region of K. servazzii was developed. The quantification was enabled using a standard curve created from serially diluted plasmids containing the target sequence of the K. servazzii strain. A validation of the assay was achieved by enumeration of K. servazzii DNA copies from artificially infected culture broths containing non-contaminated pizza substrates. The newly developed method was then tested on total DNA extracted from packaged fresh pizzas, in which certain lots were swollen and thus suspected of containing K. servazzii. This study highlights that this newly developed quantitative assay is not only sufficiently sensitive, specific and reliable to be functionally used in food control as a routine method of detection, but also promising in specific studies that seek to further characterize the dynamic of this yeast in some increasingly popular food processes.
authors.affiliation: Laboratory of Biotechnology and Applied Biology, Haute Ecole Provinciale de Hainaut-CONDORCET, 17 Chemin du Champ de Mars, 7000 Mons, Wallonia, Belgium. Electronic address: martin.spanoghe@condorcet.be.
authors.forename: Martin
authors.initials: M
authors.lastname: Spanoghe
authors.affiliation: Laboratory of Biotechnology and Applied Biology, Haute Ecole Provinciale de Hainaut-CONDORCET, 17 Chemin du Champ de Mars, 7000 Mons, Wallonia, Belgium; Centre pour l'Agronomie et l'Agro-Industrie de la Province de Hainaut (CARAH), 11 rue Paul Pastur, 7800, Ath, Wallonia, Belgium.
authors.forename: Mario
authors.initials: M
authors.lastname: Godoy Jara
authors.affiliation: Laboratory of Biotechnology and Applied Biology, Haute Ecole Provinciale de Hainaut-CONDORCET, 17 Chemin du Champ de Mars, 7000 Mons, Wallonia, Belgium.
authors.forename: John
authors.initials: J
authors.lastname: Rivière
authors.affiliation: Laboratory of Biotechnology and Applied Biology, Haute Ecole Provinciale de Hainaut-CONDORCET, 17 Chemin du Champ de Mars, 7000 Mons, Wallonia, Belgium.
authors.forename: Deborah
authors.initials: D
authors.lastname: Lanterbecq
authors.affiliation: Laboratory of Biotechnology and Applied Biology, Haute Ecole Provinciale de Hainaut-CONDORCET, 17 Chemin du Champ de Mars, 7000 Mons, Wallonia, Belgium; Centre pour l'Agronomie et l'Agro-Industrie de la Province de Hainaut (CARAH), 11 rue Paul Pastur, 7800, Ath, Wallonia, Belgium.
authors.forename: Martine
authors.initials: M
authors.lastname: Gadenne
authors.affiliation: Laboratory of Biotechnology and Applied Biology, Haute Ecole Provinciale de Hainaut-CONDORCET, 17 Chemin du Champ de Mars, 7000 Mons, Wallonia, Belgium.
authors.forename: Thierry
authors.initials: T
authors.lastname: Marique
mesh_terms: D017931:DNA Primers; D005516:Food Microbiology / Q000379:methods*; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods*; D018570:Risk Assessment; D012441:Saccharomyces cerevisiae / Q000235:genetics; D004718:Saccharomycetales / Q000235:genetics / Q000302:isolation & purification*; D013045:Species Specificity
chemical_list: D017931:DNA Primers
keywords: Food; Package swelling; Pizza; Saccharomycetaceae; Spoilage; qPCR detection