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Home / Equine Research Bank / Exp Appl Acarol / Evaluation of a mimotope of the Rickettsia outer membrane pr...
Experimental & applied acarology2023; 89(2); 317-327; doi: 10.1007/s10493-023-00776-5

Evaluation of a mimotope of the Rickettsia outer membrane protein A (OmpA) as an antigen in enzyme-linked immunosorbent assay to detect rickettsiosis in capybaras (Hydrochoerus hydrochaeris), horses (Equus caballus), and opossums (Didelphis sp.).

Abstract: Rickettsia rickettsii is the etiological agent of Rocky Mountain spotted fever, which is an important tick-borne zoonosis and, in Brazil, it causes Brazilian spotted fever, which has high lethality rate. This study aimed to evaluate a synthetic peptide corresponding to a segment of the outer membrane protein A (OmpA) as an antigen in a serological test for the diagnosis of rickettsial infections. The amino acid sequence of the peptide was selected by predicting B cell epitopes using B Cell Epitope Prediction (Immune Epitope Database and Analysis Resource) and Epitopia and OmpA sequences of Rickettsia rickettsii strain 'Brazil' and Rickettsia parkeri strains 'Maculatum 20' and 'Portsmouth'. A peptide with amino acid sequence common to both Rickettsia species was synthesized and arbitrarily named OmpA-pLMC. To evaluate this peptide in enzyme-linked immunosorbent assay (ELISA), serum samples of capybara (Hydrochoerus hydrochaeris), horse (Equus caballus), and opossum (Didelphis albiventris) that had been previously tested by indirect immunofluorescence assay (IFA) for rickettsial infection were separated into IFA-positive and IFA-negative groups and used in the assay. There were no significant differences in ELISA optical density (OD) values between IFA-positive and IFA-negative groups with horse samples. The mean OD values were significantly higher in the IFA-positive capybara serum samples (IFA-pos vs. IFA-neg = 2.389 ± 0.761 vs. 1.760 ± 0.840). However, receiver operating characteristic (ROC) curve analysis did not show significant diagnostic parameters. On the other hand, 12 out of 14 (85.7%) opossum samples of the IFA-positive group showed reactivity in ELISA, and this was significantly higher than of the IFA-negative group (0.7196 ± 0.440 vs. 0.2318 ± 0.098, respectively; 85.7% sensitivity, 100% specificity). Therefore, our results show that OmpA-pLMC has a potential to be used in immunodiagnostic assays to detect spotted fever group rickettsial infections.
© 2023. The Author(s), under exclusive licence to Springer Nature Switzerland AG.
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Publication Date: 2023-02-16 PubMed ID: 36795267PubMed Central: 6126329DOI: 10.1007/s10493-023-00776-5Google Scholar: Lookup
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Summary

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The research article presents a study aimed at assessing the effectiveness of a synthetic peptide (part of a protein) in diagnosing Rickettsia infections (like Rocky Mountain spotted fever), particularly in certain animals such as capybaras, horses, and opossums, using an enzyme-linked immunosorbent (ELISA) test.

Research Purpose and Methodology

  • The main objective of the research was to evaluate a synthetic peptide derived from a segment of the outer membrane protein A (OmpA) in rickettsia, and test its effectiveness as an antigen in serological (blood serum) tests for diagnosing rickettsial infections. This is important because Rickettsia are the causative agents of diseases like Rocky Mountain spotted fever, which have high fatality rates.
  • The researchers first selected the amino acid sequence of the peptide based on predicted B cell epitopes. This utilized the Immune Epitope Database and Analysis Resource and Epitopia, and the OmpA sequences from two Rickettsia strains.
  • The selected peptide, named OmpA-pLMC, had a sequence common to both Rickettsia species and was synthesized for the experiment.
  • To assess the effectiveness of this peptide in ELISA tests, serum samples from capybaras, horses, and opossums were separated into groups based on their prior indirect immunofluorescence assay (IFA) testing results for rickettsial infection: IFA-positive and IFA-negative.
  • The ELISA test was then conducted with the new antigen and the results compared between the two groups.

Main Findings

  • The researchers found no significant difference in ELISA results between IFA-positive and IFA-negative groups using horse serum samples.
  • However, for capybara serum samples, mean ELISA results significantly differed between the IFA-positive and IFA-negative groups. Despite this, receiver operating characteristic (ROC) curve analysis did not indicate significant diagnostic parameters.
  • In opossum samples, the IFA-positive group demonstrated significant reactivity in ELISA with the new antigen, showing much higher average results than the IFA-negative group. The results indicated an 85.7% sensitivity rate and 100% specificity rate.
  • Based on these findings, the researchers concluded that the OmpA-pLMC peptide has potential for use in immunodiagnostic assays to detect Rickettsia infections, belonging to the spotted fever group, especially in opossums.

Cite This Article

APA
Muniz APM, Tolesano-Pascoli G, Vieira RBK, Polli MG, Rodrigues VDS, Gonzaga HT, Mamede CCN, Da Cunha NC, Szabó MJP, Yokosawa J. (2023). Evaluation of a mimotope of the Rickettsia outer membrane protein A (OmpA) as an antigen in enzyme-linked immunosorbent assay to detect rickettsiosis in capybaras (Hydrochoerus hydrochaeris), horses (Equus caballus), and opossums (Didelphis sp.). Exp Appl Acarol, 89(2), 317-327. https://doi.org/10.1007/s10493-023-00776-5

Publication

Experimental & applied acarology
ISSN: 1572-9702
NlmUniqueID: 8507436
Country: Netherlands
Language: English
Volume: 89
Issue: 2
Pages: 317-327

Researcher Affiliations

Muniz, Ana Paula Mendes
  • Laboratory of Microorganisms of Cerrado (Brazilian Savannah), Department of Microbiology, Instituto De Ciências Biomédicas (ICBIM), Universidade Federal de Uberlândia (UFU), Uberlândia, Brazil.
Tolesano-Pascoli, Graziela
  • Laboratory of Ixodology, Faculdade de Medicina Veterinária, UFU, Uberlândia, Brazil.
Vieira, Raíssa Brauner Kamla
  • Laboratory of Ixodology, Faculdade de Medicina Veterinária, UFU, Uberlândia, Brazil.
  • Department of Veterinary Public Health, Universidade Federal Fluminense (UFF), Niterói, Brazil.
Polli, Mayara Garcia
  • Laboratory of Microorganisms of Cerrado (Brazilian Savannah), Department of Microbiology, Instituto De Ciências Biomédicas (ICBIM), Universidade Federal de Uberlândia (UFU), Uberlândia, Brazil.
Rodrigues, Vinícius da Silva
  • Laboratory of Ixodology, Faculdade de Medicina Veterinária, UFU, Uberlândia, Brazil.
Gonzaga, Henrique Tomaz
  • Department of Parasitology, ICBIM, UFU, Uberlândia, Brazil.
Mamede, Carla Cristine Neves
  • Molecular and Cellular Biology Laboratory, ICBIM, UFU, Uberlândia, Brazil.
Da Cunha, Nathalie Costa
  • Department of Veterinary Public Health, Universidade Federal Fluminense (UFF), Niterói, Brazil.
Szabó, Matias Juan Pablo
  • Laboratory of Ixodology, Faculdade de Medicina Veterinária, UFU, Uberlândia, Brazil.
Yokosawa, Jonny
  • Laboratory of Microorganisms of Cerrado (Brazilian Savannah), Department of Microbiology, Instituto De Ciências Biomédicas (ICBIM), Universidade Federal de Uberlândia (UFU), Uberlândia, Brazil. jonny.yokosawa@ufu.br.

MeSH Terms

  • Horses
  • Animals
  • Rodentia
  • Didelphis
  • Rickettsia
  • Rickettsia Infections / diagnosis
  • Rickettsia Infections / veterinary
  • Rocky Mountain Spotted Fever / microbiology
  • Rocky Mountain Spotted Fever / veterinary
  • Rickettsia rickettsii
  • Enzyme-Linked Immunosorbent Assay / veterinary
  • Horse Diseases / diagnosis
  • Rodent Diseases / microbiology

Grant Funding

  • 445490/ 2014-4 / Conselho Nacional de Desenvolvimento Cientu00edfico e Tecnolu00f3gico - CNPq
  • PPM 0191-16 / Fundau00e7u00e3o de Amparo u00e0 Pesquisa do Estado de Minas Gerais - FAPEMIG

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