Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2016 May 21:16:219.
doi: 10.1186/s12879-016-1552-9.

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

Dehui Yin  1 , Li Li  1 , Xiuling Song  1 , Han Li  1   2 , Juan Wang  1 , Wen Ju  1 , Xiaofeng Qu  1 , Dandan Song  1 , Yushen Liu  1 , Xiangjun Meng  1 , Hongqian Cao  1 , Weiyi Song  1 , Rizeng Meng  1   3 , Jinhua Liu  3 , Juan Li  4 , Kun Xu  5
Affiliations

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

Dehui Yin et al. BMC Infect Dis. .

Abstract

Background: In 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.

Method: To 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.

Results: For 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.

Conclusion: Our 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.

Keywords: Brucellosis; Diagnosis; Recombinant protein.

PubMed Disclaimer

Figures

Fig. 1
Fig. 1
Amino acid sequence of the recombinant protein in FAST format: “GGGS” is the linker
Fig. 2
Fig. 2
Preparation and identification of recombinant protein. a SDS-PAGE of OMP-induced expression with 0.2 mM IPTG for different times (M, marker; Lane 1, uninduced cells; Lane 2, supernatant of IPTG-induced cells for 2 h; Lane 3, deposition of IPTG-induced cells for 2 h; Lane 4, supernatant of IPTG-induced cells for 4 h; Lane 5, deposition of IPTG-induced cells for 4 h). b SDS-PAGE of OMP purification (M, marker; Lane 1, purified recombinant protein.). c Western blotting analysis of the purified recombinant protein using the anti-His-tag rabbit anti-His-tag polyclonal antibody (M, marker; Lane 1, purified recombinant protein)
Fig. 3
Fig. 3
IELISA analysis of serum samples. The analysis was performed considering positive control serum samples with culture-confirmed/serologically positive brucellosis (146 sera) and negative control sera from patients with other diseases and healthy individuals (102 sera). a Dotplot of the rOMP IELISA assay. b ROC analysis of rOMP IELISA assay results. c Dotplot of the SAT antigen IELISA assay results. d ROC analysis of SAT antigen IELISA assay results
See this image and copyright information in PMC

References

    1. Zygmunt MS, Diaz MA, Teixeira-Gomes AP, Cloeckaert A. Cloning, nucleotide sequence, and expression of the brucella melitensis sucb gene coding for an immunogenic dihydrolipoamide succinyltransferase homologous protein. Infect Immun. 2001;69:6537–40. doi: 10.1128/IAI.69.10.6537-6540.2001. - DOI - PMC - PubMed
    1. Cloeckaert A, Vizcaino N, Paquet JY, Bowden RA, Elzer PH. Major outer membrane proteins of Brucella spp.: Past, present and future. Vet Microbiol. 2002;90:229–247. doi: 10.1016/S0378-1135(02)00211-0. - DOI - PubMed
    1. Pappas G, Papadimitriou P, Akritidis N, Christou L, Tsianos EV. The new global map of human brucellosis. Lancet Infect Dis. 2006;6:91–9. doi: 10.1016/S1473-3099(06)70382-6. - DOI - PubMed
    1. Shang DQ, Xiao DL, Yin JM. Epidemiology and control of brucellosis in China. Vet Microbiol. 2002;90:165–82. doi: 10.1016/S0378-1135(02)00252-3. - DOI - PubMed
    1. Wang Y, Zhang W, Ke Y, Zhen Q, Yuan X, Zou W, Li S, Sun Y, Wang Z, Wang D, Cui B, Song H, Huang L, Chen Z. Human brucellosis, a heterogeneously distributed, delayed, and misdiagnosed disease in china. Clin Infect Dis. 2013;56(5):750–1. doi: 10.1093/cid/cis980. - DOI - PubMed

Publication types