A New Recombinant Multiepitope Chimeric Protein of Leptospira interrogans Is a Promising Marker for the Serodiagnosis of Leptospirosis
Abstract
:1. Introduction
2. Materials and Methods
2.1. Bacteria and Serum Samples
2.2. Microscopic Agglutination Test (MAT)
2.3. Composition and Design of Multiepitope Chimeric Protein
2.4. Expression and Purification of Recombinant Chimeric Protein (rChi2)
2.5. Immunoblotting Assay
2.6. ELISA for Detection of Antibody against rChi2
2.7. Immunogenicity of rChi2 in Hamster Model
2.8. Anti-rChi2 Antiserum Reactivity against Different Leptospiral Species
2.9. Ethical Statements
3. Results
3.1. Chimeric Protein Composition and Epitope Conservation among Leptospira spp.
3.2. Expression and Purification of rChi2
3.3. Detection of rChi2-Reactive IgM and IgG Antibodies in Pooled Human Serum Samples
3.4. Reactivity of rChi2 with Individual Human Serum Samples
3.5. rChi2 Is Immunogenic in Hamsters and Elicits Antibodies Capable of Recognizing Pathogenic Leptospira
4. Discussion
5. Conclusions
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Protein | Conserved | Selected | Predicted |
---|---|---|---|
Domain a | Region | Epitopes b | |
LIC10123 | FecR | 132–191 | 3 |
(91–201) | |||
LIC13059 | - | 76–150 | 3 |
LIC13050 | Big | 100–140/287–324 | 2/2 |
(67–164)/(250–343) | |||
LIC12615 | DUF3383 | 120–205 | 4 |
(270–458) | |||
LIC11636 | AIM24 | 70–155 | 5 |
(3–206) | |||
LipL32 | LipL32 | 108–138/220–272 | 2 * |
(47–243) | |||
LIC11122 | FecR | 84–155 | 3 |
(53–155) | |||
LIC11089 | - | 30–105 | 5 |
LIC10713 | Peptidase_M75 | 161–203 | 2 |
(58–351) | |||
LIC13434 | Peptidase_M43 | 374–464 | 4 |
(318–466) |
10123 | 13059 | 13050 | 12615 | 11636 | LipL32 | 11122 | 11089 | 10713 | 13434 | |
---|---|---|---|---|---|---|---|---|---|---|
L. interrogans serovar Copenhageni | 100 | 100 | 100/100 | 100 | 100 | 100/100 | 100 | 100 | 100 | 100 |
L. kirschneri Cynopteri | 100 | 96 | 100/97 | 94 | 100 | 100/100 | 100 | 99 | 98 | 99 |
L. noguchii Panama | 100 | 96 | 100/97 | 94 | 95 | 100/100 | 94 | 92 | 86 | 98 |
L. alstoni Pingchang | 93 | 91 | 98/97 | 76 | 98 | 97/98 | 90 | 83 | 85 | 92 |
L. weilii Ranarum | 92 | 89 | 98/97 | 76 | 98 | 97/98 | 88 | 82 | 78 | 90 |
L. alexanderi Manhao | 95 | 91 | 100/97 | ND | 93 | 97/96 | 89 | 84 | 82 | 92 |
L. borgpetersenii Hardjo-bovis | 93 | 89 | 100/97 | ND | 90 | 100/100 | 86 | 77 | 81 | 93 |
L. santarosai Shermani | 93 | 88 | 100/100 | 40 | 90 | 100/100 | 89 | 78 | 81 | 92 |
L. kmetyi Malaysia | 97 | 41 (c = 30) | 100/95 | 80 | 95 | 97/96 | 90 | 69 | 70 | 88 |
L. fainei Hurstbridge | 69 | 39 | 63/57 | 38 | 74 | 90/75 | 62 | 43 | 52 | 56 |
L. broomii Hurstbridge | 69 | 0 | 60/62 | 44 | 75 | 85/73 | 62 | 37 | 48 | 56 |
L. wolffi Khorat | 66 | 37 (c = 46) | 70/65 | ND | 75 | 90/70 | 62 | ND | 43 | 56 |
L. liceraciae Varillal | 65 | 0 | 64/60 | ND | 71 | 90/64 | 57 | 31 | 46 | 56 |
L. inadai Lyme | 69 | 39 | 60/60 | 44 (c = 045) | 74 | 86/73 | 62 | 40 | 48 | 57 |
L. biflexa Patoc | 50 | 0 | 63/51 | ND | ND | ND/ND | 40 | ND | ND | 50 |
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Fernandes, L.G.V.; Avelar, K.E.S.; Romero, E.C.; Heinemann, M.B.; Kirchgatter, K.; Nascimento, A.L.T.O. A New Recombinant Multiepitope Chimeric Protein of Leptospira interrogans Is a Promising Marker for the Serodiagnosis of Leptospirosis. Trop. Med. Infect. Dis. 2022, 7, 362. https://doi.org/10.3390/tropicalmed7110362
Fernandes LGV, Avelar KES, Romero EC, Heinemann MB, Kirchgatter K, Nascimento ALTO. A New Recombinant Multiepitope Chimeric Protein of Leptospira interrogans Is a Promising Marker for the Serodiagnosis of Leptospirosis. Tropical Medicine and Infectious Disease. 2022; 7(11):362. https://doi.org/10.3390/tropicalmed7110362
Chicago/Turabian StyleFernandes, Luis G. V., Kátia E. S. Avelar, Eliete C. Romero, Marcos B. Heinemann, Karin Kirchgatter, and Ana L. T. O. Nascimento. 2022. "A New Recombinant Multiepitope Chimeric Protein of Leptospira interrogans Is a Promising Marker for the Serodiagnosis of Leptospirosis" Tropical Medicine and Infectious Disease 7, no. 11: 362. https://doi.org/10.3390/tropicalmed7110362