Dot-enzyme linked immunosorbent assay as an alternative technique for the detection of bovine respiratory syncytial virus (BRSV) antibodies

Abstract

A dot-ELISA test for the detection of anti-BRSV antibodies is described. The objective of this study was the standardisation of a test as a fast, inexpensive and effective alternative to detect anti-BRSV antibodies. Its sensitivity, specificity and usefulness were compared to a commercial ELISA-kit and to the standard serum neutralisation (SN) test. The standardisation of the technique was done using nitrocellulose disks soaked with a viral sample isolated in Brazil, BRSV-25-BR. The best results were obtained when the disks were sensitised with a purified antigen at a concentration of 0.7 μg/disk and the bovine serum was diluted 1: 200. The experiment used 423 samples of bovine serum collected in the main cattle breeding centres in Brazil. The standard SN, dot-ELISA technique and commercial ELISA kits scored 67.8%, 71.8% and 72.3% of the samples as positive, respectively. When compared to the SN test, the standardised dot-ELISA and the commercial ELISA tests presented relative sensitivities of 92.3% and 91.6% and relative specificities of 71.3% and 68.4% respectively. The results demonstrated that the dot-ELISA test is adequate for the objectives proposed by this study, being easy to use and economically viable. Thus, this test represents an alternative for BRSV serological diagnosis in the substitution of SN and commercial ELISA tests, recommendable for utilisation in laboratories with few resources.