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F., Görföl, T., Kemenesi, G., Igriczi, B., Balka, G., Domán, M., Bálint, Á., Zádori, Z., & Fehér, E. (2026). Surveillance of Swine Coronaviruses in Hungarian Herds with a Newly Established Pan-Coronavirus RT-PCR System. Animals, 16(3), 358.","publicationIDType":"doi","publicationIDNumber":"10.3390/ani16030358","publicationURL":"https://www.mdpi.com/2076-2615/16/3/358"},"dsDescription":{"dsDescriptionValue":"The rapid evolution of coronaviruses (CoVs) requires researchers to develop specific yet broad-spectrum detection methods to monitor their constant genomic changes. The goal of the present study was to establish a current pan-coronavirus RT-PCR system capable of detecting a wide variety of CoVs and useful for the investigation of virus diversity and host spectrum. For optimization, one-step and two-step nested RT-PCRs with three RT enzymes were examined, amplifying a ~600 bp long product of the RNA-dependent RNA polymerase. As templates, the in vitro transcribed RNA of ten pathogenic CoVs (SARS-CoV, SARS-CoV-2, NL-63, OC43, feline CoV, porcine epidemic diarrhea virus or PEDV, transmissible gastroenteritis virus or TGEV, canine CoV, bat CoV, and infectious bronchitis virus) were applied instead of the often-used DNA standards. A limit of detection of 5–50 copies/reaction was achieved with a random hexamer-primed two-step RT-PCR and a touchdown cycling profile, representing a lower detection limit and higher specificity compared to previously published primer sets. Swine origin pooled samples (n = 121), collected from apparently healthy herds in Hungary, were tested with the novel RT-PCR system. Sequences of porcine respiratory CoV/TGEV and porcine hemagglutinating encephalomyelitis virus were identified in 24 oral fluid and nasal swab pools, demonstrating the circulation of these viruses in this country, as well as the suitability of the new PCR for their detection. The results highlighted the importance of adequate RT enzyme selection and the use of RNase inhibitors in sample preparation and conservation.","dsDescriptionDate":"2026-02-23"},"datasetContact":{"datasetContactName":"Máté, Dóra","datasetContactAffiliation":"Department of Microbiology and Infectious Diseases, University of Veterinary Medicine Budapest","datasetContactEmail":"mate.dora@univet.hu"},"journalVolumeIssue":{"journalVolume":"16","journalIssue":"3","journalPubDate":"2026-01-23"},"dateOfCollection":{"dateOfCollectionStart":"2020","dateOfCollectionEnd":"2022"},"dsDescription_hu":{"dsDescriptionValue_hu":"A koronavírusok (CoV-ok) gyors evolúciója szükségessé teszi olyan specifikus, ugyanakkor széles spektrumú kimutatási módszerek fejlesztését, amelyek alkalmasak a genom folyamatos változásainak nyomon követésére. Jelen vizsgálat célja egy korszerű, pán-koronavírus RT-PCR rendszer kialakítása volt, amely a CoV-ok széles körének detektálására alkalmas, és hatékonyan alkalmazható a vírusdiverzitás és a gazdaspektrum vizsgálatában.\n\nAz optimalizálás során egy- és kétlépéses, nested RT-PCR eljárásokat vizsgáltunk három különböző reverz transzkriptáz enzim alkalmazásával, amelyek az RNS-függő RNS-polimeráz gén ~600 bp hosszúságú szakaszát amplifikálták. Mint templátokat tíz patogén CoV (SARS-CoV, SARS-CoV-2, NL63, OC43, macska CoV, sertés járványos hasmenés vírus [PEDV], fertőző gastroenteritis vírus [TGEV], kutya CoV, denevér CoV és fertőző bronchitis vírus) in vitro transzkribált RNS-e szolgált, a gyakran alkalmazott DNS-standardok helyett.\n\nA random hexamer primerekkel végzett kétlépéses RT-PCR és touchdown ciklusprofil alkalmazásával 5–50 kópia/reakció kimutatási határt sikerült elérni, amely alacsonyabb detekciós limitet és nagyobb specificitást jelentett a korábban publikált primerkészletekhez képest.\n\nA fejlesztett RT-PCR rendszert magyarországi, klinikailag egészséges sertésállományokból származó, összevont mintákon (n = 121) teszteltük. Huszonnégy szájüregi folyadék- és orrtampon-mintapoolból a sertés eredetű respiratorikus CoV/TGEV, valamint a sertés hemagglutináló encephalomyelitis vírus szekvenciái kerültek azonosításra, igazolva e vírusok hazai cirkulációját, továbbá az új PCR-rendszer alkalmasságát azok kimutatására.\n\nEredményeink rámutattak a megfelelő reverz transzkriptáz enzim kiválasztásának, valamint az RNáz-inhibitorok minták előkészítése és tárolása során történő alkalmazásának kiemelt jelentőségére."},"keyword":[{"keywordValue":"pan-coronavirus"},{"keywordValue":"PCR"},{"keywordValue":"mammalian"},{"keywordValue":"avian"},{"keywordValue":"human"},{"keywordValue":"swine"}],"geographicCoverage":{"country":"Hungary"},"topicClassification":{"topicClassValue":"molecular diagnostics"},"author":[{"authorName":"Máté, Dóra","authorAffiliation":"Department of Microbiology and Infectious Diseases, University of Veterinary Medicine Budapest","authorIdentifierScheme":"ORCID","authorIdentifier":"0009-0001-7200-4823"},{"authorName":"Varga-Kugler, Renáta","authorAffiliation":"Ceva-Phylaxia Ltd.","authorIdentifierScheme":"ORCID","authorIdentifier":"0000-0002-0093-5418"},{"authorName":"Kaszab, Eszter","authorAffiliation":"Department of Microbiology and Infectious Diseases, University of Veterinary Medicine Budapest","authorIdentifierScheme":"ORCID","authorIdentifier":"0000-0002-5750-1872"},{"authorName":"Károlyi, Henrik Fülöp","authorAffiliation":"National Laboratory of Virology, Szentágothai Research Centre, University of Pécs","authorIdentifierScheme":"ORCID","authorIdentifier":"0009-0007-5035-7701"},{"authorName":"Görföl, Tamás","authorAffiliation":"National Laboratory of Virology, Szentágothai Research Centre, University of Pécs","authorIdentifierScheme":"ORCID","authorIdentifier":"0000-0002-1910-4024"},{"authorName":"Kemenesi, Gábor","authorAffiliation":"National Laboratory of Virology, Szentágothai Research Centre, University of Pécs","authorIdentifierScheme":"ORCID","authorIdentifier":"0000-0001-9775-3065"},{"authorName":"Igriczi, Barbara","authorAffiliation":"Department of Pathology, University of Veterinary Medicine Budapest","authorIdentifierScheme":"ORCID","authorIdentifier":"0000-0001-7372-3144"},{"authorName":"Balka, Gyula","authorAffiliation":"Department of Pathology, University of Veterinary Medicine Budapest","authorIdentifierScheme":"ORCID","authorIdentifier":"0000-0003-2742-173X"},{"authorName":"Domán, Marianna","authorAffiliation":"HUN-REN Veterinary Medical Research Institute","authorIdentifierScheme":"ORCID","authorIdentifier":"0000-0002-1823-9971"},{"authorName":"Bálint, Ádám","authorAffiliation":"Vetcontrol Ltd.","authorIdentifierScheme":"ORCID","authorIdentifier":"0000-0002-2494-1097"},{"authorName":"Zádori, Zoltán","authorAffiliation":"HUN-REN Veterinary Medical Research Institute","authorIdentifierScheme":"ScopusID","authorIdentifier":"6602902322"},{"authorName":"Fehér, Enikő","authorAffiliation":"Department of Microbiology and Infectious Diseases, University of Veterinary Medicine Budapest","authorIdentifierScheme":"ORCID","authorIdentifier":"0000-0001-7778-9116"}],"grantNumber":[{"grantNumberAgency":"National Research, Development, and Innovation Office","grantNumberValue":"FK154149"},{"grantNumberAgency":"National Research, Development, and Innovation Office","grantNumberValue":"FK137778"},{"grantNumberAgency":"National Research, Development, and Innovation Office","grantNumberValue":"RRF-2.3.1-21-2022-00001"},{"grantNumberAgency":"Ministry of Culture and Innovation of Hungary","grantNumberValue":"2024-2.1.1-EKÖP-2024-00018"}],"@id":"https://hdl.handle.net/21.15109/ARP/GH1AYK","@type":["ore:Aggregation","schema:Dataset"],"schema:version":"1.1","schema:name":"Supplementary data – Hungarian swine CoVs – Máté, 2026","schema:dateModified":"2026-02-24 14:22:02.385","schema:datePublished":"2026-02-23","schema:creativeWorkStatus":"RELEASED","schema:license":"http://creativecommons.org/licenses/by-nc/4.0","dvcore:fileTermsOfAccess":{"dvcore:fileRequestAccess":true},"schema:includedInDataCatalog":"ARP","schema:isPartOf":{"schema:name":"Respiratory Bacteriology Dataverse","@id":"https://repo.researchdata.hu/dataverse/HUN-REN_VMRI_Respiratory_Bacteriology","schema:description":"https://vmri.hu/en/temacsoportok/legzoszervi-bakteriologia\n","schema:isPartOf":{"schema:name":"Veterinary Medical Research Institute (ÁTKI)","@id":"https://repo.researchdata.hu/dataverse/vmri","schema:isPartOf":{"schema:name":"Hungarian Research Network","@id":"https://repo.researchdata.hu/dataverse/hun-ren","schema:description":"<h2>A HUN-REN Magyar Kutatási Hálózat intézményi tárolója</h2>\nA HUN-REN Magyar Kutatási Hálózat a magyar tudományos élet alappillérét jelentő kutatási hálózat. 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