Abstract
Abstract
Bovine brucellosis is a disease that significantly impacts animal production and human health. Although many sensitive diagnostic tests are used, there is still no ideal fast serological test for all epidemiological situations. In this context, we developed peptides that mimic regions of antigenic proteins of Brucella abortus and can be used in serological diagnosis.
Results
From phage display technology, we randomly selected nine clones of phage displaying peptide binders to B. abortus. These clones were sequenced and translated. After molecular docking analysis, two peptides (Ba4 and Ba9) were selected, chemically synthesized, and verified for their potential diagnostic value. By enzyme-linked immunoassay (ELISA), Ba9 showed a sensitivity of up to 97.5% to detect antibodies circulating in animals with brucellosis. We incorporated the peptide Ba9 onto a bioelectrode (graphite modified with poly-3-hydroxyphenylacetic acid). Then, direct serum detection was demonstrated by differential pulse voltammetry, micrographs, and topographic analyses in addition to the average roughness coefficient (Ra) and the value of the mean squared deviation of the roughness (Rms).
Conclusion
This work shows that the mimetic epitope of B. abortus can be useful for developing new platforms for diagnosing brucellosis. In addition, we propose a fast test based on an electrochemical sensor using graphite modified with poly-3-hydroxyphenylacetic acid.
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Availability of data and material
The datasets used and/or analysed during the current study are available from the corresponding author on reasonable request.
Abbreviations
- Ba4 Ba7 and Ba9:
-
Peptides selected
- ELISA:
-
Enzyme-linked immunosorbentassay
- Ra:
-
Average roughness coefficient
- Rms:
-
Mean squared deviation of the roughness
- PCR:
-
Polymerase chain reaction
- RBT:
-
Rose bengal test
- SAT:
-
Serum agglutination test
- CFT:
-
Complement fixation test
- PD:
-
Phage display
- PBS:
-
Phosphate-buffered saline
- BSA:
-
Bovine serum albumin
- PEG:
-
Polyethylene glycol
- HRP:
-
Horseradish peroxidase
- OPD:
-
o-phenylenediamine dihydrochloride
- HPA:
-
3-hydroxyphenylacetic acid 3
- AFM:
-
Atomic force microscopy
- DPV:
-
Differential pulse voltammetry
- AUC:
-
Area under the curve
- SOD:
-
Zn superoxide dismutase
- RCSB:
-
Research Collaboratory for Structural Bioinformation
- PDB:
-
Protein Data Bank
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Acknowledgements
The authors thank Luiz Ricardo Goulart Filho for idealizing and designing this study. Your departure left us with a vast sadness, but your brilliance and generosity reached all who had the honour to learn from you. You live within us.
Funding
This research was funded by CNPq/MAPA/SDA Nº 064/2008 and the National Institute of Science and Technology in Theranostics and Nanobiotechnology –INCT-Teranano (CNPq/CAPES/FAPEMIG, Grant # CNPq-403193/2022-2).
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LRG idealized, designed, and analysed the data and corrected the manuscript. FAAS designed and conducted the phage display, ELISA, and in silico analysis and wrote the manuscript. PTF, ERV and LMB helped with phage display and ELISA tests. ACHC and AGBM constructed and characterized the bioelectrode. VAG and AMCL were responsible for obtaining and characterizing the bovine sera. BBF analysed the data, edited the article and helped with the ELISA. AMCL idealized the project.
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The use of the sera of the animals used was allowed by the Ethics Committee on the Use of Animals of the Federal University of Uberlândia (0,99/11, 088/12, 043/14).
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Santos, F.d., Fujimura, P.T., Vaz, E.R. et al. Selection of Brucella abortus mimetic epitopes for fast diagnostic purposes in cattle. Vet Res Commun 47, 987–997 (2023). https://doi.org/10.1007/s11259-022-10043-4
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DOI: https://doi.org/10.1007/s11259-022-10043-4