Evaluation of a recombinant multiepitope antigen for diagnosis of hepatitis C virus: A lower cost alternative for antigen production

Thomasini, Ronaldo Luis; Souza, Hortencia Gisele Amaro; Bruna-Romero, Oscar; Tótola, Antônio Helvécio; Gonçales, Neiva Sellan Lopes; Lima, Cristiano Xavier; Teixeira, Mauro Martins

doi:10.1002/jcla.22410

Cited by 9 publications

(7 citation statements)

References 17 publications

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“…Several multiepitope proteins for the diagnosis of different diseases, both human [ 40 , 41 , 42 , 44 , 45 , 46 , 47 ] and animal [ 48 , 49 ], have been described and show promising results in terms of increasing and improving sensitivity and specificity and reducing cross-reactivity, indicating that multiepitope technology is efficient for the detection of different pathologies. Few studies in the literature have used recombinant multiepitope to diagnose CD.…”

Section: Discussionmentioning

confidence: 99%

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

et al. 2023

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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.

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Section: Discussionmentioning

confidence: 99%

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

et al. 2023

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“…Several multiepitope proteins for the diagnosis of different diseases, both human [42][43][44][46][47][48][49][50] and animal [51][52][53], have been described and show promising results in terms of increasing and improving sensitivity and specificity and reducing cross-reactivity, indicating that multiepitope technology is efficient for the detection of different pathologies. There are few studies in the literature using recombinant multiepitope to diagnose CD.…”

Section: Discussionmentioning

confidence: 99%

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

Machado¹,

Pereira²,

Maia³

et al. 2022

Preprint

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Background: Chagas disease is still a neglected disease considered a public health problem. 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 the problems currently faced. This study aimed to develop a new recombinant multiepitope protein for the diagnosis of Chagas disease, hereafter named rTC. Methods: The rTC was constructed based on amino acid sequences from different combinations of Trypanossoma cruzi antigens in the same polypeptide and tested in ELISA for detecting different Chagas disease. Results: rTC1 was able to discriminate between indeterminate (IND) and cardiac (CARD) cases and cross-reactive diseases, and healthy samples, with 96.6% sensitivity and 97.96% specificity, respectively. Conclusion: These data suggest a preliminary study that rTC1 has the po-tential to be tested in future studies against a larger serological panel for the diagnosis of Chagas disease.

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“…coli cells Indirect ELISA 147 foot-and-mouth-positive serum samples/610 healthy swine serum samples Sensitivity: 95.9% Specificity: 96.7% [ 105 ]/China Human hepatitis C/hepatitis C virus Recombinant multiepitope HCV antigen E . coli cells ELISA 88 hepatitis-positive serum samples/376 hepatitis C-negative serum samples/18 serum samples from non-hepatitis C diseases Sensitivity: 100% Specificity: 99.73% [ 106 ]/Brazil Human cytomegalovirus/cytomegalovirus rMEHCMV E. coli BL21(DE3) cells ELISA 12 cytomegalovirus-positive serum samples/1 serum sample from healthy individual rMEHCMV was recognized by human positive samples [ 107 ]/Brazil Canine coronavirus/SARS-CoV-2 rSP E. coli BL21(DE3) cells Indirect ELISA 64 coronavirus-positive serum samples/10 healthy dog serum samples 82.81% positive rate [ 108 ]/China Human coronavirus/SARS-CoV-2 Dx-SARS2-RBD and Dx-SARS2-noRBD E. coli BL21(DE3) cells ELISA 185 coronavirus-positive serum samples/256 serum samples from healthy individuals/94 serum samples from non-coronovirus diseases Dx-SARS2-RBD Sensitivity: 100% Specificity: 99.51-100% Dx-SARS2-noRBD Sensitivity: 100% Specificity: 99.21-100% [ 109 ]/Brazil African swine fever/African swine fever virus reMeP72 E. coli BL21(DE3) cells Colloidal gold-based immunochromatographic assay 139 swine serum samples Sensitivity: 85.7% Specificity: 97.6% [ 110 ]/China …”

Section: Rmps Applied In Disease Diagnosismentioning

confidence: 99%

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

Gonçalves,

Ribeiro,

Resende

et al. 2024

Microb Cell Fact

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Recombinant multiepitope proteins (RMPs) are a promising alternative for application in diagnostic tests and, given their wide application in the most diverse diseases, this review article aims to survey the use of these antigens for diagnosis, as well as discuss the main points surrounding these antigens. RMPs usually consisting of linear, immunodominant, and phylogenetically conserved epitopes, has been applied in the experimental diagnosis of various human and animal diseases, such as leishmaniasis, brucellosis, cysticercosis, Chagas disease, hepatitis, leptospirosis, leprosy, filariasis, schistosomiasis, dengue, and COVID-19. The synthetic genes for these epitopes are joined to code a single RMP, either with spacers or fused, with different biochemical properties. The epitopes’ high density within the RMPs contributes to a high degree of sensitivity and specificity. The RMPs can also sidestep the need for multiple peptide synthesis or multiple recombinant proteins, reducing costs and enhancing the standardization conditions for immunoassays. Methods such as bioinformatics and circular dichroism have been widely applied in the development of new RMPs, helping to guide their construction and better understand their structure. Several RMPs have been expressed, mainly using the Escherichia coli expression system, highlighting the importance of these cells in the biotechnological field. In fact, technological advances in this area, offering a wide range of different strains to be used, make these cells the most widely used expression platform. RMPs have been experimentally used to diagnose a broad range of illnesses in the laboratory, suggesting they could also be useful for accurate diagnoses commercially. On this point, the RMP method offers a tempting substitute for the production of promising antigens used to assemble commercial diagnostic kits.

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Evaluation of a recombinant multiepitope antigen for diagnosis of hepatitis C virus: A lower cost alternative for antigen production

Abstract: The multiepitope protein used here has a lower cost compared to production of each antigen separately and could be an alternative for the serological diagnosis of hepatitis C.

Cited by 9 publications

References 17 publications

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

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

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

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

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