A Multi-Epitope Protein for High-Performance Serodiagnosis of Chronic Chagas Disease in ELISA and Lateral Flow Platforms

Overview

Journal Int J Mol Sci

Publisher MDPI

Specialties Biochemistry
Chemistry
Molecular Biology

Date 2024 Sep 28

PMID 39337297

Authors

Evandro R Dias

Andressa M Durans

Barbara B Succar

Luiz Andre L T Pinto

Guilherme C Lechuga

Mariana G Miguez

Janaina Figueira-Mansur

Ana P C Argondizzo

Aline R Bernardo

Rafaela L Diniz

Gabriela S Esteves

Edimilson D Silva

Carlos M Morel

Jose Borges-Pereira

Salvatore G De-Simone

Angela C V Junqueira

David William Provance Jr

Affiliations

Soon will be listed here.

Abstract

We developed a protein to rapidly and accurately diagnose Chagas disease, a life-threatening illness identified by the WHO as a critical worldwide public health risk. Limitations in present day serological tests are complicating the current health situation and contributing to most infected persons being unaware of their condition and therefore untreated. To improve diagnostic testing, we developed an immunological mimic of the etiological agent, , by combining ten pathogen-specific epitopes within the beta-barrel protein structure of Thermal Green Protein. The resulting multi-epitope protein, DxCruziV3, displayed high specificity and sensitivity as the antibody capture reagent in an ELISA platform with an analytical sensitivity that exceeds WHO recommendations. Within an immunochromatographic platform, DxCruziV3 showed excellent performance for the point of application diagnosis in a region endemic for multiple diseases, the municipality of Barcelos in the state of Amazonas, Brazil. In total, 167 individuals were rapidly tested using whole blood from a finger stick. As recommended by the Brazilian Ministry of Health, venous blood samples were laboratory tested by conventional assays for comparison. Test results suggest utilizing DxCruziV3 in different assay platforms can confidently diagnose chronic infections by . Rapid and more accurate results will benefit everyone but will have the most noticeable impact in resource-limited rural areas where the disease is endemic.

Citing Articles

New Epitopes for the Serodiagnosis of Human Borreliosis.

Alcon-Chino M, Bonoldi V, Pereira R, Gazeta G, Carvalho J, Napoleao-Pego P Microorganisms. 2024; 12(11).

PMID: 39597601 PMC: 11596413. DOI: 10.3390/microorganisms12112212.

References

Coura J, Albajar Vinas P . Chagas disease: a new worldwide challenge. Nature. 2010; 465(7301):S6-7. DOI: 10.1038/nature09221. View

Ibanez C, Affranchino J, Macina R, Reyes M, Leguizamon S, Camargo M . Multiple Trypanosoma cruzi antigens containing tandemly repeated amino acid sequence motifs. Mol Biochem Parasitol. 1988; 30(1):27-33. DOI: 10.1016/0166-6851(88)90129-6. View

Coura J, Junqueira A . Ecological diversity of Trypanosoma cruzi transmission in the Amazon basin. The main scenaries in the Brazilian Amazon. Acta Trop. 2015; 151:51-7. DOI: 10.1016/j.actatropica.2015.04.029. View

Bradford M . A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976; 72:248-54. DOI: 10.1016/0003-2697(76)90527-3. View

Alonso-Padilla J, Abril M, Alarcon De Noya B, Almeida I, Angheben A, Araujo Jorge T . Target product profile for a test for the early assessment of treatment efficacy in Chagas disease patients: An expert consensus. PLoS Negl Trop Dis. 2020; 14(4):e0008035. PMC: 7179829. DOI: 10.1371/journal.pntd.0008035. View

Houghton R, Benson D, Reynolds L, McNEILL P, Sleath P, Lodes M . A multi-epitope synthetic peptide and recombinant protein for the detection of antibodies to Trypanosoma cruzi in radioimmunoprecipitation-confirmed and consensus-positive sera. J Infect Dis. 1999; 179(5):1226-34. DOI: 10.1086/314723. View

Zhang Y . I-TASSER server for protein 3D structure prediction. BMC Bioinformatics. 2008; 9:40. PMC: 2245901. DOI: 10.1186/1471-2105-9-40. View

Dantas R . Management of Esophageal Dysphagia in Chagas Disease. Dysphagia. 2021; 36(3):517-522. DOI: 10.1007/s00455-021-10297-1. View

Carmona-Castro O, Moo-Llanes D, Ramsey J . Impact of climate change on vector transmission of Trypanosoma cruzi (Chagas, 1909) in North America. Med Vet Entomol. 2017; 32(1):84-101. DOI: 10.1111/mve.12269. View

10.

Daltro R, Leony L, Freitas N, Silva A, Santos E, Del-Rei R . Cross-Reactivity Using Chimeric Trypanosoma cruzi Antigens: Diagnostic Performance in Settings Where Chagas Disease and American Cutaneous or Visceral Leishmaniasis Are Coendemic. J Clin Microbiol. 2019; 57(8). PMC: 6663885. DOI: 10.1128/JCM.00762-19. View

11.

Massad E . The elimination of Chagas' disease from Brazil. Epidemiol Infect. 2007; 136(9):1153-64. PMC: 2870925. DOI: 10.1017/S0950268807009879. View

12.

Ricci A, Bracco L, Salas-Sarduy E, Ramsey J, Nolan M, Lynn M . The Trypanosoma cruzi Antigen and Epitope Atlas: antibody specificities in Chagas disease patients across the Americas. Nat Commun. 2023; 14(1):1850. PMC: 10070320. DOI: 10.1038/s41467-023-37522-9. View

13.

Thomas M, Fernandez-Villegas A, Carrilero B, Maranon C, Saura D, Noya O . Characterization of an immunodominant antigenic epitope from Trypanosoma cruzi as a biomarker of chronic Chagas' disease pathology. Clin Vaccine Immunol. 2011; 19(2):167-73. PMC: 3272920. DOI: 10.1128/CVI.05566-11. View

14.

Coura J . The discovery of chagas disease (1908-1909): great successes and certain misunderstandings and challenges. Rev Soc Bras Med Trop. 2013; 46(4):389-90. DOI: 10.1590/0037-8682-0143-2013. View

15.

Peralta J, Teixeira M, Shreffler W, Pereira J, Burns Jr J, Sleath P . Serodiagnosis of Chagas' disease by enzyme-linked immunosorbent assay using two synthetic peptides as antigens. J Clin Microbiol. 1994; 32(4):971-4. PMC: 267164. DOI: 10.1128/jcm.32.4.971-974.1994. View

16.

Studier F, Moffatt B . Use of bacteriophage T7 RNA polymerase to direct selective high-level expression of cloned genes. J Mol Biol. 1986; 189(1):113-30. DOI: 10.1016/0022-2836(86)90385-2. View

17.

Thomas M, Longobardo M, Carmelo E, Maranon C, Planelles L, Patarroyo M . Mapping of the antigenic determinants of the T. cruzi kinetoplastid membrane protein-11. Identification of a linear epitope specifically recognized by human Chagasic sera. Clin Exp Immunol. 2001; 123(3):465-71. PMC: 1906009. DOI: 10.1046/j.1365-2249.2001.01478.x. View

18.

Coura J, Albajar Vinas P, Junqueira A . Ecoepidemiology, short history and control of Chagas disease in the endemic countries and the new challenge for non-endemic countries. Mem Inst Oswaldo Cruz. 2014; 109(7):856-62. PMC: 4296489. DOI: 10.1590/0074-0276140236. View

19.

Sanchez-Camargo C, Albajar-Vinas P, Wilkins P, Nieto J, Leiby D, Paris L . Comparative evaluation of 11 commercialized rapid diagnostic tests for detecting Trypanosoma cruzi antibodies in serum banks in areas of endemicity and nonendemicity. J Clin Microbiol. 2014; 52(7):2506-12. PMC: 4097696. DOI: 10.1128/JCM.00144-14. View

20.

Coura J, Dias J . Epidemiology, control and surveillance of Chagas disease: 100 years after its discovery. Mem Inst Oswaldo Cruz. 2009; 104 Suppl 1:31-40. DOI: 10.1590/s0074-02762009000900006. View