Estela Bonzo

Enzootic bovine leukosis: performance of an indirect ELISA applied in serological diagnosis

Bovine Leukosis Virus (EBLV) is a widely distributed pathogen agent in the bovine population of many countries, especially in dairy cattle. Once the bovine is infected, it remains as a virus carrier for life and such state is correlated with a specific antibody detectable level. In this study the evaluation of an indirect ELISA (Leucokit-La Plata) to detect antibodies against EBLV is presented. Comparing it with the Immunodiffusion as gold standard test, the sensitivity is 98.93%, the specificity 79.74%, the negative predictive value 99.56% and the positive predictive value 61.26%. The correspondence between both tests is 83.9% which is similar to the result mentioned by other authors (82.2%). The concordance was evaluated by calculating Kappa and Youdens J coefficients, obtaining values classified as good for both coefficients. Comparing Leucokit-La Plata and another commercial reference kit (Chekit Leucotest Bommeli AG, Bern Switzerland), the sensitivity (97.05%), specificity (94.11%), negative predictive value (92.30%) and positive predictive value (97.77%), were obtained. Applying Kappa and Joudens Index (J) coefficients an almost perfect concordance was obtained between both kits. The Leucokit-La Plata is appropriate to apply to the commercialization of live bovines to export, bovine selection for hemo-vaccines and the implementation of control and eradication programmes.

Enzootic bovine Leukosis: development of an indirect enzyme linked immunosorbent assay (I-Elisa) in seroepidemiological studies

Bovine Leukemia Virus (BLV) is the etiologic agent of Enzootic Bovine Leukosis, a retrovirus exogenous to the bovine species. Once infected, there is no detectable viraemia but instead there is a strong and persistent immunological response to BLV structural proteins, essentially the gp51 envelope glycoprotein and the mayor core protein p24. We describe the test procedure of an indirect ELISA (I-ELISA) using polyclonal reagents for the detection of antibodies to BLV. For comparison, the sera were simultaneously tested by agar gel immunodiffussion (AGID) test, which is currently used as diagnostic standard for BLV infection. The antigen applied does not require a high degree of purification and the data from the analysis of the negative sera showed that the establishment of a cut-off level corresponding to 3 times the standard deviation (SD) above the mean for the negative control set of sera provided acceptable specificity, reducing the risk of false positives results. A comparison of the results obtained by AGID test and I-ELISA showed that considering a total of 465 serum samples, all of the 234 samples (50%) that were positive by AGID were positive to the I-ELISA. Of 225 serum samples which yielded negative results in the AGID test, 69 (15%) were found to be positive by the I-ELISA and 156 (33%) were negative by both techniques. Few sera (2%) that were non-specific by AGID were defined as negative or positive by I-ELISA.

Prevalence of antibodies against Kilham virus in experimental rat colonies of Argentina

The Kilham rat virus (KRV) is a parvovirus originally isolated from a rat sarcoma in the late 1950s. The clinical signs associated with a natural KRV infection include foetal resorption in dams, runting, ataxia, cerebellar hypoplasia and jaundice in suckling rats, and sudden death, scrotal cyanosis, abdominal swelling and dehydration in juvenile rats. The ability of this virus to produce persistent infections has resulted in a high frequency of contamination of cell cultures and transplantable-tumor system. In addition, the virus may interfere with research in other ways. The remarkable resistance to environmental conditions determines the importance of the detection and control of this agent, especially in the laboratory animal production. This study determines the seroprevalence of Kilham antibodies from sera of adult rats from conventional facilities, using the haemagglutination inhibition test. The seroprevalence varied between 27.8% and 75%. This result confirms that the virus is circulating in Argentinean conventional facilities and might be interfering with research. The recognized Kilham virus may be prevented from supply sources by implementing a health monitoring schedule including a regular serological surveillance, and by keeping the animals under barrier systems.