Screening Brucella spp. in bovine raw milk by real-time quantitative PCR and conventional methods in a pilot region of vaccination, Edirne, Turkey

Basit Öğe Kaydı
dc.authorwosidŞEN, Burhan/AAF-3466-2019
dc.authorwosidOkten, Suzan/HGV-1334-2022
dc.authorwosidÖkten, Suzan/HJH-6316-2023
dc.authorwosidONURDAG, Fatma KAYNAK/T-2518-2017
dc.contributor.authorKaynak-Onurdag, F.
dc.contributor.authorOkten, S.
dc.contributor.authorSen, B.
dc.date.accessioned2024-06-12T10:55:20Z
dc.date.available2024-06-12T10:55:20Z
dc.date.issued2016
dc.departmentTrakya Üniversitesien_US
dc.description.abstractBrucellosis is a worldwide zoonotic disease transmitted to humans by consumption of contaminated milk and milk products. Brucellosis is endemic in Turkey, and Edirne has a high Brucella prevalence. Brucellosis is prevented by live-attenuated vaccines for animals and the vaccination program has been in place since 1984 in Turkey. Thrace is the pilot region for this vaccination program. The gold standard diagnostic technique for brucellosis is still the isolation of suspicious bacterial colonies followed by bacteriological identification, but it is very time consuming and laborious. In many studies, Brucella has been investigated by PCR techniques. However, PCR-based methods cannot differentiate between the vaccine strain and the virulent strain; thus, the vaccine strain may interfere with the virulent strain and causes false-positive reactions. To monitor brucellosis control programs effectively, it is important to distinguish vaccine and field strains of Brucella spp. In this study, raw milk samples were collected from 99 cows at 12 different barns in 5 villages of Edirne (Turkey). Bacteriological analyses and real-time quantitative (q)PCR experiments were applied to all samples. The DNA was isolated using Biospeedy DNA-Tricky Purification Kit (Bioeksen, Istanbul, Turkey). For all reactions, Roche Light Cycler Nano (Roche Diagnostics, Mannheim,.Germany) instrument and Biospeedy EvaGreen qPCR Pre-Mix (Bioeksen) were used.. The data were analyzed,using Roche Light-Cycler NanoSoftware 1.0. For samples that were negative by bacteriological analyses and positive by qPCR, we developed a novel qPCR-based method to differentiate the virulent B. abortus strains and B. abortus S19 vaccine strain. We designed qPCR primers targeting the outer membrane protein of B. abortus. The qPCR products were sequenced using the ABI Prism Big Dye Terminator Cycle Sequencing Ready Reaction Kit on an ABI Prism 377 DNA sequencer (Applied Biosystems, Foster City, CA). In total, 2.02% of the samples were Brucella positive, by both bacteriological method and the novel qPCR method. We concluded that, to obtain true-positive results in Brucella spp. screening studies for milk, differentiating the virulent and vaccine strain should not be disregarded.en_US
dc.description.sponsorshipTrakya University Scientific Projects Institution (Erdine, Turkey)en_US
dc.description.sponsorshipThis study was supported by the 2013-151 coded scientific research project financed by Trakya University Scientific Projects Institution (Erdine, Turkey).en_US
dc.identifier.doi10.3168/jds.2015-10637
dc.identifier.endpage3357en_US
dc.identifier.issn0022-0302
dc.identifier.issn1525-3198
dc.identifier.issue5en_US
dc.identifier.pmid26971148en_US
dc.identifier.scopus2-s2.0-84959924518en_US
dc.identifier.scopusqualityQ1en_US
dc.identifier.startpage3351en_US
dc.identifier.urihttps://doi.org/10.3168/jds.2015-10637
dc.identifier.urihttps://hdl.handle.net/20.500.14551/19385
dc.identifier.volume99en_US
dc.identifier.wosWOS:000374712100013en_US
dc.identifier.wosqualityQ1en_US
dc.indekslendigikaynakWeb of Scienceen_US
dc.indekslendigikaynakScopusen_US
dc.indekslendigikaynakPubMeden_US
dc.language.isoenen_US
dc.publisherElsevier Science Incen_US
dc.relation.ispartofJournal Of Dairy Scienceen_US
dc.relation.publicationcategoryMakale - Uluslararası Hakemli Dergi - Kurum Öğretim Elemanıen_US
dc.rightsinfo:eu-repo/semantics/openAccessen_US
dc.subjectBrucella Spp.en_US
dc.subjectMilken_US
dc.subjectQpcren_US
dc.subjectVirulent Strainen_US
dc.subjectVaccine Strainen_US
dc.subjectB. Suisen_US
dc.subjectAbortusen_US
dc.subjectStrainen_US
dc.subjectDifferentiationen_US
dc.subjectAssayen_US
dc.subjectIdentificationen_US
dc.subjectMelitensisen_US
dc.subjectSamplesen_US
dc.subjectCattleen_US
dc.subjectBlooden_US
dc.titleScreening Brucella spp. in bovine raw milk by real-time quantitative PCR and conventional methods in a pilot region of vaccination, Edirne, Turkeyen_US
dc.typeArticleen_US

Dosyalar

Koleksiyon

WoS İndeksli Yayınlar Koleksiyonu
PubMed İndeksli Yayınlar Koleksiyonu
Scopus İndeksli Yayınlar Koleksiyonu