Study on antigenic protein Omp2b in combination with Omp31 and BP26 for serological detection of human brucellosis

Brucellosis is a very common zoonosis in certain localized areas worldwide, with a high prevalence in most developing countries. The detection of brucellosis still faces many challenges such as the need for more sensitive and specific diagnostic antigens.

To evaluate the efficacy of Brucella outer membrane proteins (Omps) Omp2b in combination with omp31 and BP26 as diagnostic antigens for the serological detection of human brucellosis, these proteins were prepared by a prokaryotic expression system. Human brucellosis-positive and-negative sera were collected, and the detection effects of the diagnostic antigens were evaluated using an established indirect ELISA (iELISA) method. Receiver operating characteristic (ROC) curves and the area under the ROC curve (AUC), true positives, true negatives, false positives, false negatives, accuracy, positive predictive value, negative predictive value, analytical specificity, and sensitivity were obtained to evaluate the effectiveness of Omp2b and antigen combinations.

The iELISA results showed that the AUC of the antigenic proteins was 0.9100, 0.9387, 0.9343, and 0.9448, respectively, and that the combination of Omp31 and BP26 improved the accuracy and was superior to that of Omp2b alone. Analysis at the determined cut-off values showed that the analytical sensitivity of the assay was 0.8739 (95% CI:0.7974-0.9293) and the analytical specificity was 0.8539 (95% CI:0.7632-0.9199) when using Omp2b alone and 0.8649 when using the combination of Omp2b + BP26 (95% CI:0.7869-0.9223) with an analytical specificity of 0.9213 (95% CI:0.8446-0.9678) and 0.8468 (95% CI:0.7662-0.9082) and an analytical sensitivity of 0.9101 (95% CI:0.8305-0.9604). When Omp2b + Omp31 + BP26 was combined, the analytical sensitivity and specificity were 0.8559 (95% CI:0.7765-0.9153) and 0.9326 (95% CI:0.8590-0.9749), respectively. Protein antigens, including antigen combinations, did not cross-react with Yersinia enterocolitica O9 and E. coli O157: H7, indicating that their specificity was better than that of lipopolysaccharide (LPS).

Compared with individual Omp2b, antigen combinations improved the effectiveness in detecting brucellosis, but were still not as effective as LPS antigen. Omp2b, combined with Omp31 and BP26 as diagnostic antigens, can be used to detect human brucellosis.