Abstract"rucellosis is considered the major zoonosis in developing countries. In susceptible animal species, diagnosis of brucellosis remains a challenge due to the variety of clinical signs that it shares with a wide range of diseases. "t present, isolation of Brucella is considered the gold standard for diagnosis of brucellosis because of its low sensitivity and becoming potentially hazardous to laboratory technicians, serology is used for the detection of specific antibodies induced by infection. However, since traditional methods commonly show drawbacks and do not differentiate between vaccinated and naturally infected animals, it is necessary to search and test immunoreactive molecules for specific diagnosis of Brucella-infected cattle, thus significantly reducing the killing of suspected herds mainly due to vaccination. "dvances in biotechnology have allowed exploring the use of recombinant proteins as antigens to avoid the risk involved in the use of viable Brucella strains. The benefit of using recombinant proteins, such as outer membrane proteins OMP and other non-lipopolysaccharide non-LPS molecules as antigens, for serological diagnosis is promising, but there are still many concerns about their application. The aim of the present work is to show advances in the use of recombinant antigens and discuss their advantages and potential use as markers for the serological diagnosis in brucellosis.
The milk ring test is a detection assay for antibodies against Brucella in bovine milk. It has good sensitivity but tends to give false positive results. In this study, we standardized the application of the fluorescence polarization assay (FPA) for the detection of antibodies against B.melitensis in goat milk. We obtained negative serum and milk samples from healthy goat flocks in the northern zone of Nuevo León. Positive milk and negative, weak, and strong controls were obtained by mixing volumes of positive control serum with negative control milk. Milk samples were treated with citric acid, after which an FPA was performed. Results were then compared with the Rose Bengal test and the FPA in serum. Milk treatment allowed the quantification of antibodies in samples. Significant differences were found between the 2%, 4%, and 6% groups, compared with the control group (F3, 67 = 17.45, p < 0.0001) but not between the 2% and 4% groups (p = 0.0718). The cut-off value was 74.1 mP, with a sensitivity (Se) of 95% and a specificity (Sp) of 100%. Se and Sp values in field milk samples were 84% and 74.55%, respectively. Despite the FPA test on milk samples showed lower Se and Sp than the FPA test on serum samples, its cutoff may be adjusted. It may be recommended as a screening test in goat milk and become useful for the control and eradication of the disease.
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