Author, as appears in the article.: Salas-Masso, Nuria; Quyen Than Linh; Chin, Wai Hoe; Wolff, Anders; Andree, Karl B.; Dolors Furones, M.; Jose Figueras, Maria; Dang Duong Bang;
Department: Ciències Mèdiques Bàsiques
URV's Author/s: Figueras Salvat, María Josefa
Keywords: arcobacter campylobacter chicken carcasses escherichia-coli ethidium monoazide genetic diversity multiplex pcr pma propidium monoazide qpcr selective detection shellfish staphylococcus-aureus viable cells Arcobacter Pma Qpcr Real-time pcr Shellfish Viable cells
Abstract: The genus Arcobacter (Vandamme et al., 1991), comprised of Campylobacter-related species, are considered zoonotic emergent pathogens. The presence of Arcobacter in food products like shellfish, has an elevated incidence worldwide. In this study, we developed a specific viable quantitative PCR (v-qPCR), using the dye propidium monoazide (PMA), for quantification of the viable Arcobacter spp. cells in raw oysters and mussels. The high selectivity of primers was demonstrated by using purified DNA from 38 different species, 20 of them from the genus Arcobacter. The optimization of PMA concentration showed that 20 ?M was considered as an optimal concentration that inhibits the signal from dead cells at different concentrations (OD550 from 0.2 to 0.8) and at different ratios of live: dead cells (50:50 and 90:10). The v-qPCR results from shellfish samples were compared with those obtained in parallel using several culture isolation approaches (i.e., direct plating on marine and blood agar and by post-enrichment culturing in both media). The enrichment was performed in parallel in Arcobacter-CAT broth with and without adding NaCl. Additionally, the v-qPCR results were compared to those obtained with traditional quantitative (qPCR). The v-qPCR and the qPCR resulted in c.a. 94% of positive detection of Arcobacter vs. 41% obtained by culture approaches. When examining the reduction effect resulting from the use of v-qPCR, samples pre-enriched in Arcobacter-CAT broth supplemented with 2.5% NaCl showed a higher reduction (3.27 log copies) than that of samples obtained directly and those pre-enriched in Arcobacter-CAT broth isolation (1.05 and 1.04). When the v-qPCR was applied to detect arcobacter from real shellfish samples, 15/17 samples tested positive for viable Arcobacter with 3.41 to 8.70 log copies 1g-1. This study offers a new tool for Arcobacter surveillance in seafood.
Thematic Areas: Astronomia / física Biodiversidade Biotecnología Ciência da computação Ciência de alimentos Ciências agrárias i Ciências ambientais Ciências biológicas i Ciências biológicas ii Ciências biológicas iii Economia Engenharias i Engenharias ii Engenharias iii Ensino Farmacia Geociências Geografía Interdisciplinar Matemática / probabilidade e estatística Materiais Medicina i Medicina ii Medicina veterinaria Microbiology Microbiology (medical) Nutrição Odontología Química Saúde coletiva Zootecnia / recursos pesqueiros
licence for use: https://creativecommons.org/licenses/by/3.0/es/
Author's mail: mariajose.figueras@urv.cat
ISSN: 1664302X
Author identifier: 0000-0002-2268-8980
Record's date: 2023-02-22
Papper version: info:eu-repo/semantics/publishedVersion
Papper original source: Frontiers In Microbiology. 10 (368): 368-
APA: Salas-Masso, Nuria; Quyen Than Linh; Chin, Wai Hoe; Wolff, Anders; Andree, Karl B.; Dolors Furones, M.; Jose Figueras, Maria; Dang Duong Bang; (2019). The Use of a DNA-Intercalating Dye for Quantitative Detection of Viable Arcobacter spp. Cells (v-qPCR) in Shellfish. Frontiers In Microbiology, 10(368), 368-. DOI: 10.3389/fmicb.2019.00368
Licence document URL: https://repositori.urv.cat/ca/proteccio-de-dades/
Entity: Universitat Rovira i Virgili
Journal publication year: 2019
Publication Type: Journal Publications
Repositori URV