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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

This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.

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

References

This article includes 43 references
  1. Alves Ada S, Melo AL, Amorim MV, Borges AM, Gaíva E Silva L, Martins TF, Labruna MB, Aguiar DM, Pacheco RC. Seroprevalence of Rickettsia spp. in Equids and Molecular Detection of 'Candidatus Rickettsia amblyommii' in Amblyomma cajennense Sensu Lato Ticks From the Pantanal Region of Mato Grosso, Brazil.. J Med Entomol 2014 Nov 1;51(6):1242-7.
    doi: 10.1603/me14042pubmed: 26309313google scholar: lookup
  2. Amrun SN, Yee WX, Abu Bakar F. Novel differential linear B-cell epitopes to identify Zika and dengue virus infections in patients.. Clin Transl Immunol 8:1–15.
    doi: 10.1002/cti2.1066google scholar: lookup
  3. Avila G, Cruz-Licea V, Rojas-Espinosa K, Bermúdez-Álvarez Y, Grostieta E, Romero-Valdovinos M, Martínez-Hernández F, Vaughan G, Flisser A. Influenza A H1N1 Virus 2009 Synthetic Hemagglutinin and Neuraminidase Peptides for Antibody Detection.. Arch Med Res 2020 Jul;51(5):436-443.
    doi: 10.1016/j.arcmed.2020.04.011pubmed: 32362450google scholar: lookup
  4. Barbieri AR, Filho JM, Nieri-Bastos FA, Souza JC Jr, Szabó MP, Labruna MB. Epidemiology of Rickettsia sp. strain Atlantic rainforest in a spotted fever-endemic area of southern Brazil.. Ticks Tick Borne Dis 2014 Oct;5(6):848-53.
    doi: 10.1016/j.ttbdis.2014.07.010pubmed: 25108786google scholar: lookup
  5. Blanton LS. The rickettsioses: a practical update.. Infect Dis Clin N Am 33:213–229.
    doi: 10.1016/j.idc.2018.10.010google scholar: lookup
  6. Brazil (2021) Brazilian Ministry of Health—Boletim Epidemiológico—Doenças tropicais negligenciadas. https://www.gov.br/saude/pt-br/media/pdf/2021/marco/3/boletim_especial_doencas_negligenciadas.pdf/view . Accessed 14 March 2022
  7. Cai Y, Song Y, Cen D, Zhang C, Mao S, Ye X, Xiong Y, Jiang P, Chen J, Xue X, Zhang L, Zhu G. Novel ELISA for serodiagnosis of nasopharyngeal carcinoma based on a B cell epitope of Epstein-Barr virus latent membrane protein 2.. Oncol Lett 2018 Oct;16(4):4372-4378.
    doi: 10.3892/ol.2018.9216pubmed: 30214572pmc: 6126329google scholar: lookup
  8. Campo DS, Dimitrova Z, Yokosawa J, Hoang D, Perez NO, Ramachandran S, Khudyakov Y. Hepatitis C virus antigenic convergence.. Sci Rep 2012;2:267.
    doi: 10.1038/srep00267pubmed: 22355779pmc: 3279735google scholar: lookup
  9. Clements ML, Dumler JS, Fiset P, Wisseman CL Jr, Snyder MJ, Levine MM. Serodiagnosis of Rocky Mountain spotted fever: comparison of IgM and IgG enzyme-linked immunosorbent assays and indirect fluorescent antibody test.. J Infect Dis 1983 Nov;148(5):876-80.
    doi: 10.1093/infdis/148.5.876pubmed: 6415180google scholar: lookup
  10. Coelho MG, Ramos Vdo N, Limongi JE, de Lemos ER, Guterres A, da Costa Neto SF, Rozental T, Bonvicino CR, D'Andrea PS, Moraes-Filho J, Labruna MB, Szabó MP. Serologic evidence of the exposure of small mammals to spotted-fever Rickettsia and Rickettsia bellii in Minas Gerais, Brazil.. J Infect Dev Ctries 2016 Mar 31;10(3):275-82.
    doi: 10.3855/jidc.7084pubmed: 27031460google scholar: lookup
  11. D'Auria SR, Camargo MC, Pacheco RC, Savani ES, Dias MA, da Rosa AR, de Almeida MF, Labruna MB. Serologic survey for rickettsiosis in bats from São Paulo city, Brazil.. Vector Borne Zoonotic Dis 2010 Jun;10(5):459-63.
    doi: 10.1089/vbz.2009.0070pubmed: 19877815google scholar: lookup
  12. de Oliveira EJ, Kanamura HY, Takei K, Hirata RD, Valli LC, Nguyen NY, de Carvalho Rodrigues I, de Jesus AR, Hirata MH. Synthetic peptides as an antigenic base in an ELISA for laboratory diagnosis of schistosomiasis mansoni.. Trans R Soc Trop Med Hyg 2008 Apr;102(4):360-6.
    doi: 10.1016/j.trstmh.2007.11.008pubmed: 18314149google scholar: lookup
  13. Do EJ, Kim JE, Park JM, Lee KM, Jung MY, Lee HJ, Cho HW, Choi YJ, Lee SH, Park KH, Jang WJ. Development of recombinant OmpA and OmpB proteins as diagnostic antigens for rickettsial disease.. Microbiol Immunol 2009 Jul;53(7):368-74.
    doi: 10.1111/j.1348-0421.2009.00142.xpubmed: 19563395google scholar: lookup
  14. Fang R, Blanton LS, Walker DH. Rickettsiae as Emerging Infectious Agents.. Clin Lab Med 2017 Jun;37(2):383-400.
    doi: 10.1016/j.cll.2017.01.009pubmed: 28457356google scholar: lookup
  15. Fortes FS, Santos LC, Cubas ZS. Anti-Rickettsia spp. antibodies in free-ranging and captive capybaras from southern Brazil.. Pesqui Vet Bras 31:1014–1018.
    doi: 10.1590/S0100-736X2011001100013google scholar: lookup
  16. Freire MCLC, Pol-Fachin L, Coêlho DF, Viana IFT, Magalhães T, Cordeiro MT, Fischer N, Loeffler FF, Jaenisch T, Franca RF, Marques ETA, Lins RD. Mapping Putative B-Cell Zika Virus NS1 Epitopes Provides Molecular Basis for Anti-NS1 Antibody Discrimination between Zika and Dengue Viruses.. ACS Omega 2017 Jul 31;2(7):3913-3920.
    doi: 10.1021/acsomega.7b00608pubmed: 30023708pmc: 6044859google scholar: lookup
  17. Gottlieb M, Long B, Koyfman A. The Evaluation and Management of Rocky Mountain Spotted Fever in the Emergency Department: a Review of the Literature.. J Emerg Med 2018 Jul;55(1):42-50.
    doi: 10.1016/j.jemermed.2018.02.043pubmed: 29685474google scholar: lookup
  18. Guevarra LA Jr, Boado KJO, Ceñidoza FBB, Imbao MRLM, Sia MJG, Dalmacio LMM. A synthetic peptide analog of in silico-predicted immunogenic epitope unique to dengue virus serotype 2 NS1 antigen specifically binds immunoglobulin G antibodies raised in rabbits.. Microbiol Immunol 2020 Feb;64(2):153-161.
    doi: 10.1111/1348-0421.12757pubmed: 31710119google scholar: lookup
  19. Horta MC, Labruna MB, Sangioni LA, Vianna MC, Gennari SM, Galvão MA, Mafra CL, Vidotto O, Schumaker TT, Walker DH. Prevalence of antibodies to spotted fever group rickettsiae in humans and domestic animals in a Brazilian spotted fever-endemic area in the state of São Paulo, Brazil: serologic evidence for infection by Rickettsia rickettsii and another spotted fever group Rickettsia.. Am J Trop Med Hyg 2004 Jul;71(1):93-7.
    doi: 10.4269/ajtmh.2004.71.93pubmed: 15238696google scholar: lookup
  20. Jeannin P, Magistrelli G, Goetsch L. Outer membrane protein A (OmpA): a new pathogen-associated molecular pattern that interacts with antigen presenting cells—impact on vaccine strategies.. Vaccine 20:23–27.
    doi: 10.1016/S0264-410X(02)00383-3google scholar: lookup
  21. La Scola B, Raoult D. Laboratory diagnosis of rickettsioses: current approaches to diagnosis of old and new rickettsial diseases.. J Clin Microbiol 1997 Nov;35(11):2715-27.
    doi: 10.1128/jcm.35.11.2715-2727.1997pubmed: 9350721pmc: 230049google scholar: lookup
  22. Labruna MB. Ecology of rickettsia in South America.. Ann N Y Acad Sci 2009 May;1166:156-66.
    doi: 10.1111/j.1749-6632.2009.04516.xpubmed: 19538276google scholar: lookup
  23. Labruna MB, Horta MC, Aguiar DM, Cavalcante GT, Pinter A, Gennari SM, Camargo LM. Prevalence of Rickettsia infection in dogs from the urban and rural areas of Monte Negro municipality, western Amazon, Brazil.. Vector Borne Zoonotic Dis 2007 Summer;7(2):249-55.
    doi: 10.1089/vbz.2006.0621pubmed: 17627445google scholar: lookup
  24. Mazigi O, Schofield P, Langley DB, Christ D. Protein A superantigen: structure, engineering and molecular basis of antibody recognition.. Protein Eng Des Sel 2019 Dec 31;32(8):359-366.
    doi: 10.1093/protein/gzz026pubmed: 31641749google scholar: lookup
  25. McFee RB. Tick borne illness - Rocky mountain spotted fever.. Dis Mon 2018 May;64(5):185-194.
    doi: 10.1016/j.disamonth.2018.01.006pubmed: 29549965google scholar: lookup
  26. Milagres BS, Padilha AF, Barcelos RM, Gomes GG, Montandon CE, Pena DC, Nieri Bastos FA, Silveira I, Pacheco R, Labruna MB, Bouyer DH, Freitas RN, Walker DH, Mafra CL, Galvao MA. Rickettsia in synanthropic and domestic animals and their hosts from two areas of low endemicity for Brazilian spotted fever in the eastern region of Minas Gerais, Brazil.. Am J Trop Med Hyg 2010 Dec;83(6):1305-7.
    doi: 10.4269/ajtmh.2010.10-0239pubmed: 21118939pmc: 2990049google scholar: lookup
  27. Neves LC, Barreto ALG, de Souza MX. Serosurvey on rickettsiae of the spotted fever group and Rickettsia bellii among dogs in the state of Goiás, Brazil.. Rev Bras Parasitol Vet 29:1–5.
    doi: 10.1590/S1984-29612020018google scholar: lookup
  28. Noriea NF, Clark TR, Hackstadt T. Targeted knockout of the Rickettsia rickettsii OmpA surface antigen does not diminish virulence in a mammalian model system.. mbio 6:1–9.
    doi: 10.1128/mBio.00323-15google scholar: lookup
  29. Noriea NF, Clark TR, Mead D, Hackstadt T. Proteolytic cleavage of the immunodominant outer membrane protein rOmpA in Rickettsia rickettsii.. J Bacteriol 199:1–13.
    doi: 10.1128/jb.00826-16google scholar: lookup
  30. Paddock CD, Denison AM, Lash RR, Liu L, Bollweg BC, Dahlgren FS, Kanamura CT, Angerami RN, Pereira dos Santos FC, Brasil Martines R, Karpathy SE. Phylogeography of Rickettsia rickettsii genotypes associated with fatal Rocky Mountain spotted fever.. Am J Trop Med Hyg 2014 Sep;91(3):589-97.
    doi: 10.4269/ajtmh.14-0146pubmed: 24957541pmc: 4155566google scholar: lookup
  31. Paddock CD, Finley RW, Wright CS, Robinson HN, Schrodt BJ, Lane CC, Ekenna O, Blass MA, Tamminga CL, Ohl CA, McLellan SL, Goddard J, Holman RC, Openshaw JJ, Sumner JW, Zaki SR, Eremeeva ME. Rickettsia parkeri rickettsiosis and its clinical distinction from Rocky Mountain spotted fever.. Clin Infect Dis 2008 Nov 1;47(9):1188-96.
    doi: 10.1086/592254pubmed: 18808353google scholar: lookup
  32. Paris DH, Dumler JS. State of the art of diagnosis of rickettsial diseases: the use of blood specimens for diagnosis of scrub typhus, spotted fever group rickettsiosis, and murine typhus.. Curr Opin Infect Dis 2016 Oct;29(5):433-9.
    doi: 10.1097/QCO.0000000000000298pubmed: 27429138pmc: 5029442google scholar: lookup
  33. Potocnakova L, Bhide M, Pulzova LB. An introduction to B-cell epitope mapping and in silico epitope prediction.. J Immunol Res 2016:1–11.
    doi: 10.1155/2016/6760830google scholar: lookup
  34. Prado IC, Chino META, Dos Santos AL, Souza ALA, Pinho LG, Lemos ERS, De-Simone SG. Development of an electrochemical immunosensor for the diagnostic testing of spotted fever using synthetic peptides.. Biosens Bioelectron 2018 Feb 15;100:115-121.
    doi: 10.1016/j.bios.2017.08.029pubmed: 28886455google scholar: lookup
  35. Ramírez-Hernández A, Uchoa F, de Azevedo Serpa MC. Clinical and serological evaluation of capybaras (Hydrochoerus hydrochaeris) successively exposed to an Amblyomma sculptum-derived strain of Rickettsia rickettsii.. Sci Rep 10:1–15.
    doi: 10.1038/s41598-020-57607-5google scholar: lookup
  36. Ramírez-Hernández A, Uchoa F, Serpa MCA, Binder LC, Souza CE, Labruna MB. Capybaras (Hydrochoerus hydrochaeris) as amplifying hosts of Rickettsia rickettsii to Amblyomma sculptum ticks: Evaluation during primary and subsequent exposures to R. rickettsii infection.. Ticks Tick Borne Dis 2020 Sep;11(5):101463.
    doi: 10.1016/j.ttbdis.2020b.101463pubmed: 32723630google scholar: lookup
  37. Rigi G, Ghaedmohammadi S, Ahmadian G. A comprehensive review on staphylococcal protein A (SpA): Its production and applications.. Biotechnol Appl Biochem 2019 May;66(3):454-464.
    doi: 10.1002/bab.1742pubmed: 30869160google scholar: lookup
  38. Rodrigues-da-Silva RN, Correa-Moreira D, Soares IF, de-Luca PM, Totino PRR, Morgado FN, Oliveira Henriques MDG, Peixoto Candea AL, Singh B, Galinski MR, Moreno A, Oliveira-Ferreira J, Lima-Junior JDC. Immunogenicity of synthetic peptide constructs based on PvMSP9(E795-A808), a linear B-cell epitope of the P. vivax Merozoite Surface Protein-9.. Vaccine 2019 Jan 7;37(2):306-313.
    doi: 10.1016/j.vaccine.2018.10.016pubmed: 30509693google scholar: lookup
  39. Souza CE, Camargo LB, Pinter A, Donalisio MR. High seroprevalence for Rickettsia rickettsii in equines suggests risk of human infection in silent areas for the Brazilian spotted fever.. PLoS ONE 11:11–19.
    doi: 10.1371/journal.pone.0153303google scholar: lookup
  40. Szabó MPJ, Pinter A, Labruna MB. Ecology, biology and distribution of spotted-fever tick vectors in Brazil.. Front Cell Infect Microbiol 3:1–9.
    doi: 10.3389/fcimb.2013.00027google scholar: lookup
  41. Tolesano-Pascoli GV. Ticks and Rickettsiae in an urban park in Uberlândia, Minas Gerais: associations with ecology and biodiversity.. .
  42. Ueno TEH, Costa FB, Moraes-Filho J. Experimental infection of horses with Rickettsia rickettsii.. Parasit Vectors 9:1–11.
    doi: 10.1186/s13071-016-1784-ygoogle scholar: lookup
  43. Vieira RBK. Pesquisa de rickettsias em capivaras de áreas antropizadas, equinos de carroceiros e carrapatos de Uberlândia-MG e Itaperuna-RJ, Brasil.. .

Citations

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