Recently, a radioimmunoassay (RIA) for measurement of canine pancreatic lipase immunoreactivity (cPLI) in serum was developed and validated. However, RIAs require frequent use of radioactive materials. Therefore, the goal of this project was to develop and validate an enzyme-linked immunosorbent assay (ELISA) for cPLI. After purifying cPL, we developed and purified antiserum against cPL in rabbits. The purified antibody was bound to microtitre plates and used to capture antigen. A portion of the purified antibody was biotinylated and used to identify the captured antigen. Streptavidin labelled with horseradish peroxidase and a horseradish peroxidase substrate were used for detection. The assay was validated by determination of sensitivity, working range, linearity, accuracy, precision, and reproducibility. The reference interval for serum cPLI was determined by the central 95th percentile in 74 clinically healthy dogs: 2.2 to 102.1 μg/L. The sensitivity and the upper limit of the working range were 0.1 and 999.2 μg/L, respectively. The ratios of observed to expected values for dilutional parallelism for 6 serum samples ranged from 0.0 to 148.8%; the ratios for spiking recovery for 4 serum samples ranged from 90.4 to 112.6%, assuming 55% recovery of the cPL. Coefficients of variation for intra- and interassay variability for 6 different serum samples were 2.4, 3.4, 4.1, 5.8, 7.4, and 10.0% and 5.9, 7.7, 11.6, 13.9, 23.5, and 46.2%, respectively. We conclude that the ELISA described here is sufficiently sensitive, linear, accurate, precise, and reproducible for clinical application. Evaluation of its clinical usefulness for the diagnosis of exocrine pancreatic disorders in dogs is under way.

Six tests for the detection of West Nile virus (WNV) antibodies in the serum of experimentally infected chickens were compared. The tests included the hemagglutination-inhibition test (HIT), immunoglobulin M (IgM)-capture enzyme-linked immunosorbent assay (ELISA) with WNV-infected mouse brain antigen, immunoglobulin G (IgG) indirect ELISA with tickborne encephalitis viral antigen, the microtitre virus neutralization test, the standard plaque reduction neutralization test (PRNT), and the microtitre PRNT (micro-PRNT). Thirty adult chickens, intravenously and intramuscularly inoculated with 107 plaque-forming units (PFU) of WNV strain Egypt 101, were bled and given a booster of 107 PFU at 7, 15, and 21 d postinoculation; the final blood collection was on day 28. Although the micro-PRNT is capable of detecting the highest antibody titres during both early and late infection, because of the technical complexity and time requirements of this test a combination of IgM and IgG ELISAs is recommended for serologic screening. Serum samples that give positive results in the ELISAs can then be tested by the micro-PRNT to determine the specificity of antibodies to WNV.

An inactivated oil-based Newcastle disease vaccine was prepared using Mukteswar vaccine strain. The virus was propagated in 10-day old embryonating eggs and inactivated by 0.12% formalin for 48 hours at 37 degree C. The vaccine was formulated with 1 part antigen (aqueous phase) and 4 parts oil base. The oil base contained Tween-80 1%, Arlacel-A 10% and Mineral oil 89%. The stability of the vaccine was found satisfactory after 6 months and its viscosity and injectability was fairly ideal. The antigenicity of the vaccine was determined in 16 week-old pullets. The seromonitoring of the vaccinated and the control pullets was carried out for three months post- vaccination by Haemagglutination inhibition (HI) test. Blood samples were taken at fortnightly intervals. The Geometric mean HI titre of the vaccinated pullets on the day of vaccination 15, 30, 45, 60, 75, 90, 105 and 120 days post- vaccination was 18.4, 4.9, 87.5, 192.3, 257.6, 111.4, 91.7, 63.9 and 30.0. However, in non-vaccinated control pullets it was found to be 18.4, 3.7, 3.7, 4.3, 3.8, 4.0, 3.5, 2.8 and 2.3 respectively. The inactivated oil-based vaccine induced a marked antibody response which continued upto three months.