A novel multi-epitope recombined protein for diagnosis of human brucellosis

Abstract

BackgroundIn epidemic regions of the world, brucellosis is a reemerging zoonosis with minimal mortality but is a serious public hygiene problem. Currently, there are various methods for brucellosis diagnosis, however few of them are available to be used to diagnose, especially for serious cross-reaction with other bacteria.MethodTo overcome this disadvantage, we explored a novel multi-epitope recombinant protein as human brucellosis diagnostic antigen. We established an indirect enzyme-linked immunosorbent assay (ELISA) based on this recombinant protein. 248 sera obtained from three different groups including patients with brucellosis (146 samples), non-brucellosis patients (82 samples), and healthy individuals (20 samples) were tested by indirect ELISA. To evaluate the assay, a receiver-operating characteristic (ROC) analysis and immunoblotting were carried out using these characterized serum samples.ResultsFor this test, the area under the ROC curve was 0.9409 (95 % confidence interval, 0.9108 to 0.9709), and a sensitivity of 88.89 % and a specificity of 85.54 % was given with a cutoff value of 0.3865 from this ROC analysis. The Western blot results indicate that it is feasible to differentiate human brucellosis and non-brucellosis with the newly established method based on this recombinant protein.ConclusionOur results obtained high diagnostic accuracy of the ELISA assay which encourage the use of this novel recombinant protein as diagnostic antigen to implement serological diagnosis of brucellosis.Electronic supplementary materialThe online version of this article (doi:10.1186/s12879-016-1552-9) contains supplementary material, which is available to authorized users.

Structural Basis and Designing of Peptide Vaccine using PE-PGRS Family Protein of Mycobacterium ulcerans – An Integrated Vaccinomics Approach

“…Furthermore, DNA vaccine encoding immunodominant Ag85A protein of M. ulcerans showed protection similar to BCG vaccination (Tanghe et al 2008;Hart, Hale, and Lee 2016). These evidences are suggesting that the subunit vaccine could be an alternative in terms of safety, efficacy and specificity (Saadi, Karkhah, and Nouri 2017;Yin et al 2016).…”

“…However, the development of effective multi-epitope vaccines remains challenging due to the difficulties in the selection of appropriate antigen candidates, immunodominant epitopes and an efficient delivery system (L. Zhang 2018). Therefore, the prediction of suitable antigenic epitopes of a target protein by immunoinformatic methods is extremely important for designing a multi-epitope vaccine (Yin et al 2016;Cherryholmes, Stanton, and Disis 2015).…”

A sandwich immunoassay for brucellosis diagnosis based on immune magnetic beads and quantum dots

“…In addition, the diagnostic sensitivity was 96.15% (95% CI, 91.82-98.58%) and the diagnostic specificity was 94.12% (95% CI, 87.64-97.81%) with the (Table 2). In our previous study [23], we used rOMP as the detection antigen coated into ninety-six-well microtiter plates and diagnosed the 248 serum samples by indirect ELISA. The AUC of the ROC analysis was 0.9409 (95% CI, 0.9108-0.9709).…”

“…The rOMP was prepared as described in our previous studies [23]. The predicted B cell epitopes were selected to construct the rOMP and were joined together by the linker "GGGS".…”

“…We use the rOMP as an antigen for the diagnosis of brucellosis by indirect ELISA method because the specificity and sensitivity is higher than whole cell antigen [23]. In our present study, we developed a novel brucellosis diagnostic method using the rOMP prepared in our previous studies based on nano-detection technology.…”