New Epitopes for the Serodiagnosis of Human Borreliosis
Lyme disease, a zoonotic infection caused by the bacterium <i>Borrelia burgdorferi</i>, is transmitted to humans through the bites of infected ticks. Its diagnosis primarily relies on serological methods; however, the existing borreliosis techniques have shown a variable sensitivity and...
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2024-10-01
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| author | Mônica E. T. Alcón-Chino Virgínia L. N. Bonoldi Rosa M. R. Pereira Gilberto S. Gazeta João P. R. S. Carvalho Paloma Napoleão-Pêgo Andressa M. Durans André L. A. Souza Salvatore G. De-Simone |
| author_facet | Mônica E. T. Alcón-Chino Virgínia L. N. Bonoldi Rosa M. R. Pereira Gilberto S. Gazeta João P. R. S. Carvalho Paloma Napoleão-Pêgo Andressa M. Durans André L. A. Souza Salvatore G. De-Simone |
| author_sort | Mônica E. T. Alcón-Chino |
| collection | DOAJ |
| description | Lyme disease, a zoonotic infection caused by the bacterium <i>Borrelia burgdorferi</i>, is transmitted to humans through the bites of infected ticks. Its diagnosis primarily relies on serological methods; however, the existing borreliosis techniques have shown a variable sensitivity and specificity. Our study aimed to map IgG epitopes from five outer membrane proteins (Omp) from <i>B. burgdorferi</i> [Filament flagellar 41kD (PI1089), flagellar hook-associated protein (Q44767), Flagellar hook k2 protein (O51173), Putative Omp BURGA03 (Q44849), and 31 kDa OspA (P0CL66)] lipoprotein to find specific epitopes for the development of accurate diagnosis methods. Using the spot synthesis technique, a library of 380 peptides was constructed to identify linear B cell epitopes recognized by human IgG in response to specific <i>B. burgdorferi</i>-associated proteins. The reactivity of this epitope when chemically synthesized was then evaluated using ELISA with a panel of the patient’s sera. Cross-reactivity was assessed through data bank access and in vitro analysis. Among the 19 epitopes identified, four were selected for further investigation based on their signal intensity, secondary structure, and peptide matching. Validation was performed using ELISA, and ROC curve analysis demonstrated a sensitivity of ≥85.71%, specificity of ≥92.31, accuracy of ≥90.7, and AUC value of ≥0.91 for all peptides. Our cross-reactivity analysis demonstrated that the Burg/02/huG, Burg/03/huG, and Burg/12/huG peptides were not reactive to antibodies from patients with Leptospirosis and syphilis compared to those from the <i>B. burgdorferi</i> group. These peptides indicated an excellent performance in distinguishing between <i>B. burgdorferi</i>-infected and non-infected individuals and exhibited a neglected reactivity to antibodies in sera from patients with Leptospirosis and syphilis. These peptides are promising targets for recombinant development, potentially leading to more accurate serological tests and vaccines. |
| format | Article |
| id | doaj-art-5a01065317fd4e1d81456b5c9d3e0090 |
| institution | Kabale University |
| issn | 2076-2607 |
| language | English |
| publishDate | 2024-10-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Microorganisms |
| spelling | doaj-art-5a01065317fd4e1d81456b5c9d3e00902024-11-26T18:14:32ZengMDPI AGMicroorganisms2076-26072024-10-011211221210.3390/microorganisms12112212New Epitopes for the Serodiagnosis of Human BorreliosisMônica E. T. Alcón-Chino0Virgínia L. N. Bonoldi1Rosa M. R. Pereira2Gilberto S. Gazeta3João P. R. S. Carvalho4Paloma Napoleão-Pêgo5Andressa M. Durans6André L. A. Souza7Salvatore G. De-Simone8Center for Technological Development in Health (CDTS), National Institute of Science and Technology for Innovation in Neglected Population Diseases (INCT-IDPN), FIOCRUZ, Rio de Janeiro 21040-900, RJ, BrazilClinical Hospital, Faculty of Medicine, São Paulo University, São Paulo 05403-000, SP, BrazilFaculty of Medicine, São Paulo University, São Paulo 01246-903, SP, BrazilLaboratory of Ticks and Other Wingless Arthropods-National Reference for Vectors of Rickettsioses, Instituto Oswaldo Cruz-IOC, FIOCRUZ, Rio de Janeiro 21041-250, RJ, BrazilCenter for Technological Development in Health (CDTS), National Institute of Science and Technology for Innovation in Neglected Population Diseases (INCT-IDPN), FIOCRUZ, Rio de Janeiro 21040-900, RJ, BrazilCenter for Technological Development in Health (CDTS), National Institute of Science and Technology for Innovation in Neglected Population Diseases (INCT-IDPN), FIOCRUZ, Rio de Janeiro 21040-900, RJ, BrazilCenter for Technological Development in Health (CDTS), National Institute of Science and Technology for Innovation in Neglected Population Diseases (INCT-IDPN), FIOCRUZ, Rio de Janeiro 21040-900, RJ, BrazilMultidisciplinary Biochemistry Teaching Laboratory, UNIG, Nova Iguaçu 26260-045, RJ, BrazilCenter for Technological Development in Health (CDTS), National Institute of Science and Technology for Innovation in Neglected Population Diseases (INCT-IDPN), FIOCRUZ, Rio de Janeiro 21040-900, RJ, BrazilLyme disease, a zoonotic infection caused by the bacterium <i>Borrelia burgdorferi</i>, is transmitted to humans through the bites of infected ticks. Its diagnosis primarily relies on serological methods; however, the existing borreliosis techniques have shown a variable sensitivity and specificity. Our study aimed to map IgG epitopes from five outer membrane proteins (Omp) from <i>B. burgdorferi</i> [Filament flagellar 41kD (PI1089), flagellar hook-associated protein (Q44767), Flagellar hook k2 protein (O51173), Putative Omp BURGA03 (Q44849), and 31 kDa OspA (P0CL66)] lipoprotein to find specific epitopes for the development of accurate diagnosis methods. Using the spot synthesis technique, a library of 380 peptides was constructed to identify linear B cell epitopes recognized by human IgG in response to specific <i>B. burgdorferi</i>-associated proteins. The reactivity of this epitope when chemically synthesized was then evaluated using ELISA with a panel of the patient’s sera. Cross-reactivity was assessed through data bank access and in vitro analysis. Among the 19 epitopes identified, four were selected for further investigation based on their signal intensity, secondary structure, and peptide matching. Validation was performed using ELISA, and ROC curve analysis demonstrated a sensitivity of ≥85.71%, specificity of ≥92.31, accuracy of ≥90.7, and AUC value of ≥0.91 for all peptides. Our cross-reactivity analysis demonstrated that the Burg/02/huG, Burg/03/huG, and Burg/12/huG peptides were not reactive to antibodies from patients with Leptospirosis and syphilis compared to those from the <i>B. burgdorferi</i> group. These peptides indicated an excellent performance in distinguishing between <i>B. burgdorferi</i>-infected and non-infected individuals and exhibited a neglected reactivity to antibodies in sera from patients with Leptospirosis and syphilis. These peptides are promising targets for recombinant development, potentially leading to more accurate serological tests and vaccines.https://www.mdpi.com/2076-2607/12/11/2212epitope mappingBrazilian borreliosisELISAcross-reactivity |
| spellingShingle | Mônica E. T. Alcón-Chino Virgínia L. N. Bonoldi Rosa M. R. Pereira Gilberto S. Gazeta João P. R. S. Carvalho Paloma Napoleão-Pêgo Andressa M. Durans André L. A. Souza Salvatore G. De-Simone New Epitopes for the Serodiagnosis of Human Borreliosis Microorganisms epitope mapping Brazilian borreliosis ELISA cross-reactivity |
| title | New Epitopes for the Serodiagnosis of Human Borreliosis |
| title_full | New Epitopes for the Serodiagnosis of Human Borreliosis |
| title_fullStr | New Epitopes for the Serodiagnosis of Human Borreliosis |
| title_full_unstemmed | New Epitopes for the Serodiagnosis of Human Borreliosis |
| title_short | New Epitopes for the Serodiagnosis of Human Borreliosis |
| title_sort | new epitopes for the serodiagnosis of human borreliosis |
| topic | epitope mapping Brazilian borreliosis ELISA cross-reactivity |
| url | https://www.mdpi.com/2076-2607/12/11/2212 |
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