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Testicular processing fluid as a useful matrix for the detection of porcine circovirus type 2 DNA and virus-specific antibodies

cris.virtual.author-orcid0000-0002-4509-8852
cris.virtual.author-orcid0000-0003-0270-2914
cris.virtual.author-orcid0000-0003-2220-2730
cris.virtualsource.author-orcid702c3b7e-b378-411f-bc9b-510fe73f43da
cris.virtualsource.author-orcidabb21f0b-d43c-470d-99e6-7b59a4439244
cris.virtualsource.author-orcid71dccebf-e765-40b9-87bb-e98ab3b7299c
dc.abstract.enIntroduction: Testicular processing fluid (PF) obtained during boar castration may serve as a diagnostic matrix for monitoring porcine circovirus type 2 (PCV2) presence.Methods: Anti- PCV2 antibodies in PF were detected using an indirect ELISA, and PCV2 DNA was detected by real-time PCR (qPCR), and the effects of sample pooling were evaluated. Paired sera and PF from boars and sera from gilts were tested with commercial ELISA and qPCR kits. PF-specific cut-offs were set by ROC (ELISA OD; qPCR Ct). Pooling was simulated by diluting positive PF samples with negative PF samples at predefined ratios.Results: With manufacturers’ cut-offs, seropositivity was 89.94% (male sera), 87.43% (PF), and 92.81% (gilt sera); differences were observed only between gilt sera and PF. Using the ROC PF cut-off (OD ≥ 0.23), 88.82% PF were positive, and matrices did not differ; diagnostic agreement metrics for PF improved. In qPCR, positivity was 13.02% (boar sera), 16.90% (PF), and 9.36% (gilt sera). A ROC-specific PCR cut-off (Ct < 36.50) improved specificity, predictive values, and agreement without affecting sensitivity; serum and PF Ct values were moderately correlated (ρ = 0.53). Pooling reduced detection of weak positives (most notably in qPCR) while high-positive PF remained detectable at higher dilutions.Discussion and conclusion: These results demonstrate that PF provides a reliable, cost effective alternative to serum for PCV2 surveillance and monitoring when matrix-specific cut-offs are used; however, excessive pooling may lead to false-negative results. This approach may facilitate large-scale herd monitoring while reducing the need for invasive sampling.
dc.affiliationWydział Medycyny Weterynaryjnej i Nauk o Zwierzętach
dc.affiliation.instituteKatedra Nauk Przedklinicznych i Chorób Zakaźnych
dc.contributor.authorTurlewicz-Podbielska, Hanna
dc.contributor.authorDors, Arkadiusz
dc.contributor.authorPomorska-Mól, Małgorzata
dc.date.access2026-02-23
dc.date.accessioned2026-02-23T09:06:18Z
dc.date.available2026-02-23T09:06:18Z
dc.date.copyright2026-02-19
dc.date.issued2026
dc.description.abstract<jats:sec> <jats:title>Introduction</jats:title> <jats:p>Testicular processing fluid (PF) obtained during boar castration may serve as a diagnostic matrix for monitoring porcine circovirus type 2 (PCV2) presence.</jats:p> </jats:sec> <jats:sec> <jats:title>Methods</jats:title> <jats:p>Anti- PCV2 antibodies in PF were detected using an indirect ELISA, and PCV2 DNA was detected by real-time PCR (qPCR), and the effects of sample pooling were evaluated. Paired sera and PF from boars and sera from gilts were tested with commercial ELISA and qPCR kits. PF-specific cut-offs were set by ROC (ELISA OD; qPCR Ct). Pooling was simulated by diluting positive PF samples with negative PF samples at predefined ratios.</jats:p> </jats:sec> <jats:sec> <jats:title>Results</jats:title> <jats:p> With manufacturers’ cut-offs, seropositivity was 89.94% (male sera), 87.43% (PF), and 92.81% (gilt sera); differences were observed only between gilt sera and PF. Using the ROC PF cut-off (OD ≥ 0.23), 88.82% PF were positive, and matrices did not differ; diagnostic agreement metrics for PF improved. In qPCR, positivity was 13.02% (boar sera), 16.90% (PF), and 9.36% (gilt sera). A ROC-specific PCR cut-off (Ct &amp;lt; 36.50) improved specificity, predictive values, and agreement without affecting sensitivity; serum and PF Ct values were moderately correlated ( <jats:italic>ρ</jats:italic>  = 0.53). Pooling reduced detection of weak positives (most notably in qPCR) while high-positive PF remained detectable at higher dilutions. </jats:p> </jats:sec> <jats:sec> <jats:title>Discussion and conclusion</jats:title> <jats:p>These results demonstrate that PF provides a reliable, cost effective alternative to serum for PCV2 surveillance and monitoring when matrix-specific cut-offs are used; however, excessive pooling may lead to false-negative results. This approach may facilitate large-scale herd monitoring while reducing the need for invasive sampling.</jats:p> </jats:sec>
dc.description.accesstimeat_publication
dc.description.bibliographyil., bibliogr.
dc.description.financepublication_research
dc.description.financecost14000,00
dc.description.if2,9
dc.description.points70
dc.description.versionfinal_published
dc.description.volume13
dc.identifier.doi10.3389/fvets.2026.1745725
dc.identifier.issn2297-1769
dc.identifier.urihttps://sciencerep.up.poznan.pl/handle/item/7420
dc.identifier.weblinkhttps://www.frontiersin.org/journals/veterinary-science/articles/10.3389/fvets.2026.1745725/full
dc.languageen
dc.pbn.affiliationveterinary science
dc.relation.ispartofFrontiers in Veterinary Science
dc.relation.pagesart. 1745725.
dc.rightsCC-BY
dc.sciencecloudnosend
dc.share.typeOPEN_JOURNAL
dc.subject.enlaboratory diagnostics
dc.subject.ennon-invasive matrices
dc.subject.enPCV2
dc.subject.enpig welfare
dc.subject.entesticular processing fluid
dc.titleTesticular processing fluid as a useful matrix for the detection of porcine circovirus type 2 DNA and virus-specific antibodies
dc.typeJournalArticle
dspace.entity.typePublication
oaire.citation.volume13
project.funder.nameMNiSW PREIDUB 2024-2026