A Multi-Epitope Protein for High-Performance Serodiagnosis of Chronic Chagas Disease in ELISA and Lateral Flow Platforms
Abstract
:1. Introduction
2. Results
2.1. Design and Production of DxCruziV3
2.2. Performance of DxCruziV3 in ELISAs
2.3. Analytical and Geographical Sensitivity of DxCruziV3
2.4. Lateral Flow Immunochromatographic Prototypes
2.5. Performance of the LFA Prototypes
3. Discussion
4. Materials and Methods
4.1. Ethical Approval
4.2. Serological Panels and Participant Profile
4.3. Cloning, Expression, and Purification of DxCruziV3
4.4. Enzyme-Linked Immunosorbent Assays
4.5. Lateral Flow Immunochromatographic Assay Preparation
4.6. Indirect Immunofluorescence
4.7. Statistical Analysis
5. Conclusions
6. Patents
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Epitope | Sequence | Insertion Site 1 | Color 2 | Protein Origin | Reference |
---|---|---|---|---|---|
1 | KFAELLEQQKNAQFPGK | N-term | Red | KMP11 | [26] |
2 | DSSAHSTPSTPA | 50 | Blue | SAPA | [27] |
3 | GDKPSPFGQAAAADK | 114 | Yellow | PEP-2 | [28,29] |
4 | FGQAAAGDKPS | 127 | Magenta | TcCA-2 | [30] |
5 | AEPKPAEPKS | 153 | Lt. Blue | TcD-2 | [31,32] |
6 | TSSTPPSGTENKPAT | 167 | Cyan | TSSA | [28,33] |
7 | GTSEEGSRGGSSMPS | 183 | Salmon | TcLo1.2 | [32] |
8 | SPFGQAAAGDK | 244 | Pink | B13 | [29,34] |
9 | KAAIAPA | C-term | Purple | TcE | [32] |
10 | KQRAAETK | C-term | Orange | CRA | [35] |
Assay | Repetitions 1 | Volume | Reactive | Non-Reactive |
---|---|---|---|---|
V3ib-LFA Field 2 | 1 | 10 µL | 43 | 124 |
sV3-LFA Field 2 | 1 | 10 µL | 43 | 124 |
V3ib-LFA Lab 3 | 0–1 | 5 µL | 45 | 25 |
sV3-LFA Lab (Lot#1) 3 | 0–1 | 5 µL | 44 | 123 |
sV3-LFA Lab (Lot #2) 4 | 1 | 5 µL | 36 | 131 |
V3ib ELISA (in-house) | 3–8 | 0.5–2 µL | 39 | 128 |
ELISA Wiener | 2–3 | 10 µL | 32 | 135 |
ELISA Bioclin | 2 | 10 µL | 61 | 106 |
Immunofluorescence Indirect | 1–2 | 5 µL | 48 | 119 |
Assay | Non-Divergent (n = 150) | Final Call (n = 167) | ||
---|---|---|---|---|
Sensitivity | Specificity | Sensitivity | Specificity | |
Field V3ib LFA | 100% | 100% | 95.2% | 97.6% |
Field sV3 LFA | 100% | 100% | 90.5% | 96.0% |
V3ib-ELISA | 87.5% | 98.3% | 87.5% | 97.6% |
ELISA Wiener | 71.9% | 97.5% | 64.3% | 96.0% |
ELISA Bioclin | 90.6% | 79.7% | 83.3% | 79.2% |
Immunofluorescence | 93.8% | 88.1% | 76.2% | 87.2% |
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Dias, E.R.; Durans, A.M.; Succar, B.B.; Pinto, L.A.L.T.; Lechuga, G.C.; Miguez, M.G.; Figueira-Mansur, J.; Argondizzo, A.P.C.; Bernardo, A.R.; Diniz, R.L.; et al. A Multi-Epitope Protein for High-Performance Serodiagnosis of Chronic Chagas Disease in ELISA and Lateral Flow Platforms. Int. J. Mol. Sci. 2024, 25, 9811. https://doi.org/10.3390/ijms25189811
Dias ER, Durans AM, Succar BB, Pinto LALT, Lechuga GC, Miguez MG, Figueira-Mansur J, Argondizzo APC, Bernardo AR, Diniz RL, et al. A Multi-Epitope Protein for High-Performance Serodiagnosis of Chronic Chagas Disease in ELISA and Lateral Flow Platforms. International Journal of Molecular Sciences. 2024; 25(18):9811. https://doi.org/10.3390/ijms25189811
Chicago/Turabian StyleDias, Evandro R., Andressa M. Durans, Barbara B. Succar, Luiz André L. T. Pinto, Guilherme C. Lechuga, Mariana G. Miguez, Janaina Figueira-Mansur, Ana P. C. Argondizzo, Aline R. Bernardo, Rafaela L. Diniz, and et al. 2024. "A Multi-Epitope Protein for High-Performance Serodiagnosis of Chronic Chagas Disease in ELISA and Lateral Flow Platforms" International Journal of Molecular Sciences 25, no. 18: 9811. https://doi.org/10.3390/ijms25189811