title: Development and preliminary validation of an antibody filtration-assisted single-dilution chemiluminometric immunoassay for potency testing of Piscirickettsia salmonis vaccines.
abstract: Challenge with live pathogens could be substituted by serology for many veterinary diseases, however little progress has been made in the development of alternative batch vaccine potency tests for fish. This study reports the development and preliminary validation of a single-dilution filtration-assisted chemiluminometric immunoassay (SD FAL-ELISA) applied to measure anti Piscirickettsia salmonis IgM in individual or pooled serum and mucus samples. The assay was set up to test a single-dilution of the sample. Serum SD FAL-ELISA yielded a sensitivity of 90% and a specificity of 96%. SD FAL-ELISA was applied to evaluate pooled and individual samples from P. salmonis challenge assessments. Relative-light units values (RLU) obtained by SD FAL-ELISA were proportional to antibody levels in serum. RLU values obtained from pooled and individual serum samples increased with the observed relative percent survival (RPS) values, indicating a correlation between protection and specific IgM levels. Results obtained for specific IgM in mucus samples was not related to the RPS, but discriminated the vaccine that yielded high RPS (86.4%) from the others (40.9 and 54.5%). This is the first report on the development of an indirect high-throughput serological assessment for P. salmonis vaccine potency testing using both pooled or individual serum and cutaneous mucus samples.
authors.affiliation: Instituto de Ciencia y Tecnología Dr. César Milstein, Saladillo 2468, 1440 Ciudad de Buenos Aires, Argentina.
authors.forename: Maximiliano
authors.initials: M
authors.lastname: Wilda
authors.forename: María Ángeles
authors.initials: MÁ
authors.lastname: Lavoria
authors.forename: Adrián
authors.initials: A
authors.lastname: Giráldez
authors.forename: Olga Lucía
authors.initials: OL
authors.lastname: Franco-Mahecha
authors.forename: Florencia
authors.initials: F
authors.lastname: Mansilla
authors.forename: Matías
authors.initials: M
authors.lastname: Érguiz
authors.forename: Marcela Elvira
authors.initials: ME
authors.lastname: Iglesias
authors.forename: Alejandra Victoria
authors.initials: AV
authors.lastname: Capozzo
mesh_terms: D000818:Animals; D001428:Bacterial Vaccines / Q000276:immunology*; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D049449:Luminescence; D053522:Piscirickettsia / Q000276:immunology*; D015203:Reproducibility of Results
chemical_list: D001428:Bacterial Vaccines