DEVELOPMENT OF AN ANTIBODY FILTRATION-ASSISTED SINGLE-DILUTION CHEMILUMINOMETRIC IMMUNOASSAY FOR POTENCY TESTING OF PISCIRICKETTSIA SALMONIS VACCINES

LAVORIA, MARÍA DE LOS ÁNGELES; WILDA, MAXIMILIANO; ERGUIS, MATÍAS; FRANCO MAHECHA, OLGA; GRIGERA, PABLO; IGLESIAS, MARCELA; CAPOZZO, A.V.

Lima

Congreso; InmunoPerú 2012; 2012

Asociación Latinoamericana de Inmunología

Refinement of techniques has been applied for many veterinary diseases for which challenge with live pathogens could be substituted by serology However, little progress has been made in the development of alternative batch potency tests for fish. This study reports the development of a Single-dilution Filtration-Assisted Chemiluminometric Immunoassay (SdFAL-ELISA), an easy to standardize and reliable method applied to the measurement of anti Piscirickettsia salmonis IgM in individual or pooled, serum and epithelial mucus samples. Relative-light units values (RLU) obtained by Sd-FAL-ELISA were proportional to antibody levels in serum. The assay was set up to test a single 1:200 dilution of sera or a 1:50 dilution of mucus. Sd-FAL-ELISA discriminated positive and negative samples. 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.