Validation of an indirect ELISA to detect antibodies against BoHV-1 in bovine and guinea-pig serum samples using ISO/IEC 17025 standards

Two ELISAs to quantify antibodies to BoHV-1 in the sera of cattle and immunized guinea pigs were developed and validated using ISO/IEC 17025 standards. The cut-off value of the assay was established at 20% positivity of a high positive control for screening of cattle. Using this threshold, the assay...

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Bibliographic Details
Published in:Journal of virological methods 2010-10, Vol.169 (1), p.143-153
Main Authors: Parreño, Viviana, Romera, S. Alejandra, Makek, Lucia, Rodriguez, Daniela, Malacari, Darío, Maidana, Silvina, Compaired, Diego, Combessies, Gustavo, Vena, María Marta, Garaicoechea, Lorena, Wigdorovitz, Andrés, Marangunich, Laura, Fernandez, Fernando
Format: Article
Language:English
Subjects:
Animal models
Animals
Antibodies, Viral - blood
Assay validation
Biological and medical sciences
BoHV-1
Bovine herpesvirus 1
Cattle
ELISA
Enzyme-Linked Immunosorbent Assay - methods
Enzyme-Linked Immunosorbent Assay - standards
Fundamental and applied biological sciences. Psychology
Guinea Pigs
Herpesvirus 1, Bovine - immunology
Herpesvirus Vaccines - immunology
ISO 17025
Microbiology
Neutralization Tests
Sensitivity and Specificity
Techniques used in virology
Vaccine potency testing
Virology
Virology - methods
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Summary:Two ELISAs to quantify antibodies to BoHV-1 in the sera of cattle and immunized guinea pigs were developed and validated using ISO/IEC 17025 standards. The cut-off value of the assay was established at 20% positivity of a high positive control for screening of cattle. Using this threshold, the assay properly classified the OIE bovine reference sera EU1, EU2 and EU3. For vaccine potency testing, a cut-off of 40% was selected for both species. The reliability of the assays, given by their diagnostic sensitivity and specificity, using the threshold of 40% was 89.7% and 100%, respectively, for bovines and 94.9% and 100% for guinea pigs, respectively. There was almost perfect agreement between the ELISA and virus neutralization results. In addition, after vaccination, there was a good correlation between the neutralizing and ELISA antibody titers of the serum from the same bovine or guinea pig, sampled at 60 and 30 days post-vaccination, respectively (Rbovine=0.88, Rguinea pig=0.92; p
ISSN:0166-0934
1879-0984
DOI:10.1016/j.jviromet.2010.07.014