Proof of Concept of a Novel Multiepitope Recombinant Protein for the Serodiagnosis of Patients with Chagas Disease

Abstract

Chagas disease remains a neglected disease that is considered to be a public health problem. The early diagnosis of cases is important to improve the prognosis of infected patients and prevent transmission. Serological tests are the method of choice for diagnosis. However, two serological tests are currently recommended to confirm positive cases. In this sense, more sensitive and specific serological tests need to be developed to overcome these current diagnosis problems. This study aimed to develop a new recombinant multiepitope protein for the diagnosis of Chagas disease, hereafter named rTC. The rTC was constructed based on amino acid sequences from different combinations of Trypanosoma cruzi antigens in the same polypeptide and tested using an enzyme-linked immunosorbent assay (ELISA) to detect different types of Chagas disease. rTC was able to discriminate between indeterminate (IND) and cardiac (CARD) cases and cross-reactive diseases, as well as healthy samples, with 98.28% sensitivity and 96.67% specificity, respectively. These data suggest that rTC has the potential to be tested in future studies against a larger serological panel for the diagnosis of Chagas disease.

Recombinant multiepitope proteins expressed in Escherichia coli cells and their potential for immunodiagnosis

“…infantum diseases Human and canine diagnosis Sensitivity: 100% Specificity: 100% [ 4 ]/Brazil Human Chagas disease/ Trypanosoma cruzi rTC E . coli BL21(λDE3) pLysS cells ELISA 58 Chagas-positive serum samples/30 serum samples from healthy individuals/60 serum samples from non-Chagas diseases Sensitivity: 98.28% Specificity: 96.67% [ 5 ]/Brazil …”

“…Moreover, rMELEISH demonstrated a better diagnostic performance as compared to results using soluble Leishmania antigen extract. Lastly, a novel RMP for diagnosing Chagas disease was developed by Machado et al (2023) [ 5 ]. To compose rTC antigen, the epitopes were selected based on the literature.…”

“…4 Biophysics Laboratory, Institute of Biological Sciences, Department of Cell Biology, University of Brasilia, Brasília 70910-900, Brazil. 5 Bioactives and Nanobiotechnology Laboratory, Federal University of São João Del-Rei, Midwest Campus, Divinópolis 35501-296, Brazil. 6…”

“…A study by Dipti et al (2006) is thought to be the first one to define an RMP as a molecule that contains linear, immunodominant, and conserved epitopes that are connected through linkers [1]. Since then, the commercial use of these molecules has gained market space with many applications related to human and animal health, such as the development of vaccines and diagnostic devices [3][4][5][6][7]. In fact, the global market of recombinant proteins is expected to grow by 12% from 2022 to 2030, with 2030 revenue estimated at USD 5.09 billion [8].…”