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Gewählte Publikation:

Publikationstyp: Zeitschriftenaufsatz
Dokumenttyp: Originalarbeit

Jahr: 2018

AutorInnen: Feichtner, F; Schachner, A; Berger, E; Hess, M

Titel: Fiber-based fluorescent microsphere immunoassay (FMIA) as a novel multiplex serodiagnostic tool for simultaneous detection and differentiation of all clinically relevant fowl adenovirus (FAdV) serotypes.

Quelle: J Immunol Methods. 2018; 458:33-43



Autor/innen der Vetmeduni Vienna:

Feichtner Franziska
Hess Michael
Schachner Anna

Beteiligte Vetmed-Organisationseinheiten
Universitätsklinik für Geflügel und Fische, Klinische Abteilung für Geflügelmedizin


Abstract:
The recent emergence of fowl aviadenovirus (FAdV) induced disease outbreaks in chicken flocks worldwide, with distinct aetiologies confined to particular FAdV species and serotypes, is increasingly urging the need for specific and mass-applicable antibody screening systems. Despite this exigency, there are to date no available serological procedures which satisfactorily combine the criteria for sensitive detection of antibodies against FAdVs, diagnostic reliability in face of cross-reactions and requirements for a rapid and large-scale application. In order to address this gap, a multiplexed fluorescent microsphere immunoassay (FMIA) based on recombinant FAdV fiber proteins from six different serotypes FAdV-1, -2, -4, -8a, -8b and -11 was developed, which enabled simultaneous detection of antibodies against all clinically relevant serotypes in a single reaction within a high throughput setting. Based on a panel of >300 monospecific antisera raised against each of the 12 FAdV serotypes, 100% serotype-specificity was demonstrated for FAdV-1 (FAdV-A) and FAdV-4 (FAdV-C) fiber-based analytes. Analytes based on serotypes affiliated to FAdV-D and FAdV-E exhibited moderately lower specificities of 91.2-95.7%. This was attributed almost exclusively to mutual recognition between FAdV-2 and -11 field strains and to a much lesser extent to reference strains, supporting earlier proposals to merge them into a single serotype. Similarly, extensive cross-reactions between FAdV-8a and -8b were noted. Altogether intraspecies cross-reactions can be attributed to viruses with a close etiological intersection. Antisera against other important avian viruses remained negative by the FMIA, further validating its specificity. Compared to the virus-neutralization (VN) test, FMIA and individual fiber-based enzyme-linked immunosorbent assays (ELISAs) were equally sensitive in the detection of sera against FAdV-2 and -11, as well as FAdV-8a and -8b field strains, while they were even superior to VN test in detection of FAdV-1 and FAdV-4 responses, likely attributed to a relative abundance of fiber antibodies early upon infection. Moreover, application of the FMIA on field samples comprising a diversified response against all 12 FAdV serotypes further consolidated its specificity and agreement with VN test.Copyright © 2018. Published by Elsevier B.V.

Keywords Pubmed: Animals
Antibodies, Viralbloodisolation & purification
Antigens, Viralimmunology
Aviadenovirusimmunologyisolation & purification
Capsid Proteinsimmunology
Cell Culture Techniques
Chickensvirology
Enzyme-Linked Immunosorbent Assayinstrumentationmethods
Fluoroimmunoassayinstrumentationmethods
High-Throughput Screening Assaysinstrumentationmethods
Microspheres
Poultry Diseasesblooddiagnosisvirology
Recombinant Proteinsimmunology
Reproducibility of Results
Sensitivity and Specificity
Serogroup
Serologic Testsmethods
Serotypinginstrumentationmethods
Specific Pathogen-Free Organisms

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