Evaluation of the Fluorescence Polarization Assay (FPA) for diagnosis of brucella melitensis infection of goats in Argentina.

AIM: Determine the optimal cut-off and the use of Fluorescence Polarization Assay (FPA) to improve the detection of brucellosis in individual goats in Argentina. METHODS: Sera from 96 goats from a flock with abortion due to B. melitensis biovar 1 were used to assess the efficacy of the FPA to detect brucellosis in goats. FPA results were compared with those of the Buffered Antigen Plate Agglutination test (BPAT) confirmed by Seroagglutination in tube (SAT), the competitive enzyme-linked immunosorbent assay (c-ELISA) and the indirect enzyme-linked immunosorbent assay (i-ELISA). Sera from 554 goats free from brucellosis were tested with the BPAT, SAT, c-ELISA and i-ELISA to determine its Specificity. Vaccination had not been performed in the flocks evaluated. RESULTS: The most appropriate cut-off was selected for the FPA by using MedCalc software. It was fixed at 87 mP giving a sensitivity and specificity of 98.1% (CI 89.9-99.7) and 92.8% (CI 90.4-94.7). The relative sensitivity compared with i-ELISA and c-ELISA was 97% and 92.9% respectively. The relative specificity compared with i-ELISA and c-ELISA was 97.5% and 98% respectively. The kappa measures of agreement between tests was higher than 0.75 CONCLUSION: The high correlation between FPA results and other serological methods with sera goats is indicative of the excellent performance of FPA technique in diagnosis of caprine brucellosis and we endorse it as a recommended method.

Evaluation of standard reagents for radial-immunodiffusion assays. In vitro control of rabies vaccines.

The RID assay is one of the in vitro methods used for in-process control in the production of rabies vaccines for veterinary use. It has been shown to be very useful for determining antigen concentration in the final bulk product. The work presented in this paper, including the production and standardization of candidate standard reagents for use in the Radial Immunodiffusion Assay (RID) was carried out at the Pan American Institute for Food Protection and Zoonoses (INPPAZ/PAHO/WHO). The study was completed with the cooperation of the Faculty of Veterinary Sciences, National University of La Plata (NULP), Argentina, where the validation of the proposed standards and the quality control of samples from 28 different batches of rabies vaccines produced with Pasteur strain rabies virus (PV) in BHK cells were performed. The activity of the vaccines was determined by in vivo (NIH) and in vitro (RID)assays. The results of the candidate reagents for the reagent standardization tests showed stability, sensitivity and reproducibility. The Relative Potency the 1.2 between the problem vaccines and the reference vaccine was estimated by variance and regression analysis. The results of our validation study show that the INPPAZ (PAHO/WHO) is capable of producing and distributing the above-mentioned standard reagents, as well as of providing support for the incorporation of the RID technique (sensitive, rapid and inexpensive) to the laboratories that manufacture rabies vaccines in Latin America and the Caribbean.