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Coordinated implementation of a conventional PCR assay to detect all Ebola and Marburg virus species in a European laboratory network

2025-05-28Artigo científico Acesso aberto
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datacite.subject.sdg17:Parcerias para a Implementação dos Objetivos
dc.contributor.authorHeimsch, K.C.
dc.contributor.authorBleicker, T.
dc.contributor.authorBest, T.D.
dc.contributor.authorPresser, L.D.
dc.contributor.authorMolenkamp, R.
dc.contributor.authorJääskeläinen, A.J.
dc.contributor.authorMilewska, A.
dc.contributor.authorSmahelová, J.
dc.contributor.authorBaronti, C.
dc.contributor.authorPappa, S.
dc.contributor.authorTabain, I.
dc.contributor.authorCordeiro, Rita
dc.contributor.authorMarsili, G.
dc.contributor.authorHuik, K.
dc.contributor.authordos Reis, V. Pinho
dc.contributor.authorBarzon, L.
dc.contributor.authorMaes, P.
dc.contributor.authorDrosten, C.
dc.contributor.authorCorman, V.M.
dc.date.accessioned2026-02-05T15:07:22Z
dc.date.available2026-02-05T15:07:22Z
dc.date.issued2025-05-28
dc.description.abstractBackground: Filoviruses, including Ebola and Marburg viruses, cause severe hemorrhagic fever in humans and primates. These viruses pose significant threats to public health, making rapid and sensitive detection critical for controlling outbreaks. We developed and validated a hemi-nested generic PanFilo assay to detect all Ebola virus species, Marburg viruses, and recently discovered bat filoviruses. This assay was deployed to 15 European laboratories and evaluated through testing of eight non-infectious samples. Objectives: Laboratories were asked to determine the detection limit of positive controls and test all samples using the assay provided. The deployed assay enables direct Nanopore sequencing of PCR products, by using tagged primers during the second round of PCR. Sequencing of the samples was carried out on a voluntary basis. Results: Multicenter validation revealed a 95 % limit of detection of 5309 RNA copies/μL for Ebola, 10,273 copies/μL for Marburg, and 2145 copies/μL for Mengla virus. In an implementation quality assessment, 93.3 % (84/90) of samples containing filovirus RNA were correctly identified and 100 % (30/30) of filovirus-negative samples were correctly identified. Thirteen laboratories sequenced PCR products, with nine identifying all positive samples correctly. Conclusion: The assay enables rapid and reliable detection of filoviruses, with sequencing capabilities for identifying both known and novel variants. This assay might be used for detection during the initial phase of an emerging filovirus outbreak, before a specific assay has been developed. However, our distribution across 15.eng
dc.description.abstractHighlights: A generic PanFilo PCR was deployed to 15 laboratories across 12 European countries within weeks. -The assay enables direct Nanopore sequencing of PCR products. -Successful deployment of the assay was evaluated through testing of blinded samples. -Eleven laboratories correctly identified all blinded samples. -Variability in laboratory performance highlights the need for better training and standardization.eng
dc.description.sponsorshipThis study was funded by the DURABLE project, co-funded by the European Union, under the EU4Health Programme (EU4H), project no. 101102733.
dc.identifier.citationMulticenter Study J Clin Virol. 2025 Jun:178:105808. doi: 10.1016/j.jcv.2025.105808. Epub 2025 May 28
dc.identifier.doi10.1016/j.jcv.2025.105808
dc.identifier.eissn1873-5967
dc.identifier.issn1386-6532
dc.identifier.pmid40451100
dc.identifier.urihttp://hdl.handle.net/10400.18/10817
dc.language.isoeng
dc.peerreviewedyes
dc.publisherElservier
dc.relation101102733
dc.relation.hasversionhttps://www.sciencedirect.com/science/article/pii/S1386653225000502?via%3Dihub
dc.rights.urihttp://creativecommons.org/licenses/by-nc/4.0/
dc.subjectDiagnostic Testing
dc.subjectFilovirus
dc.subjectNested PCR
dc.subjectQuality Improvement
dc.subjectInfecções Sistémicas e Zoonoses
dc.titleCoordinated implementation of a conventional PCR assay to detect all Ebola and Marburg virus species in a European laboratory networkeng
dc.typejournal article
dcterms.referenceshttps://ars.els-cdn.com/content/image/1-s2.0-S1386653225000502-mmc1.docx
dspace.entity.typePublication
oaire.citation.startPage05808
oaire.citation.titleJournal of Clinical Virology
oaire.citation.volume178
oaire.versionhttp://purl.org/coar/version/c_970fb48d4fbd8a85
person.familyNameCordeiro
person.givenNameRita
person.identifier.ciencia-idB518-A367-B8B7
person.identifier.orcid0000-0001-5321-3657
relation.isAuthorOfPublicationd94520e0-9d10-4302-af83-58afc79f152a
relation.isAuthorOfPublication.latestForDiscoveryd94520e0-9d10-4302-af83-58afc79f152a

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