Padronização de um teste Elisa indireto no diagnóstico da Cinomose em amostras de soro canino, marsupias e primatas

Abstract

Distemper is a disease caused by the Canine Distemper Virus (CDV). This disease mainly affects dogs, but also affects other domestic and wild species. The animal's immunity is related to the degree to which this pathogen will reach the individual's organism. It affects the animal's breathing, can cause vomiting, diarrhea, convulsions, and can lead to death. The objective of the present work was to standardize an indirect ELISA test to diagnose distemper in serum samples from dogs using viral surface structures as antigen. To standardize the technique, it was necessary to study the antigen dilution to identify the best concentration for plaque sensitization. The antigen was made from the isolate of the Snyder Hill strain (ATCC VR-526 – Manassas, USA) of the distemper virus and was used as an antigen for immunodiagnostic tests. The test was first applied with different dilutions of the antigen to detect the best performance of the antigen. Once this was done, it was tested in a serum bank with 90 samples, 45 of which were clinically and serologically proven negative animal samples and 45 clinically and serologically proven positive animal samples in a commercial ELISA test. A color gradient was established for reading the results without the need for a microplate reader. The cut-off point, specificity and sensitivity of the test were calculated. The indirect ELISA test proved to be excellent as a diagnostic test for canine distemper, with an optical density cutoff of 0.229, sensitivity of 95.5% and specificity of 84.4%. The created color gradient proved to be effective for reading the test results without the need for a microplate reader.