In Silico Screening of Putative Corynebacterium pseudotuberculosis Antigens and Serological Diagnosis for Caseous Lymphadenitis in Sheep by Enzyme-Linked Immunosorbent Assay
Corynebacterium pseudotuberculosis is the etiologic agent of Caseous Lymphadenitis (CLA), a disease leading to severe damage in sheep and goats farming due to the lack of serological diagnosis, treatment, and effective prophylaxis. In this context, several strategies in an attempt to discover new an...
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| Format: | Article |
| Language: | English |
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Wiley
2021-01-01
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| Series: | Veterinary Medicine International |
| Online Access: | http://dx.doi.org/10.1155/2021/9931731 |
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| author | Daniela Droppa-Almeida Caroline de Santana Ferreira Ioná Brito Sibele Borsuk Jorge Alberto López Rodríguez Isabel Bezerra Lima-Verde Francine Ferreira Padilha |
| author_facet | Daniela Droppa-Almeida Caroline de Santana Ferreira Ioná Brito Sibele Borsuk Jorge Alberto López Rodríguez Isabel Bezerra Lima-Verde Francine Ferreira Padilha |
| author_sort | Daniela Droppa-Almeida |
| collection | DOAJ |
| description | Corynebacterium pseudotuberculosis is the etiologic agent of Caseous Lymphadenitis (CLA), a disease leading to severe damage in sheep and goats farming due to the lack of serological diagnosis, treatment, and effective prophylaxis. In this context, several strategies in an attempt to discover new antigens to compose diagnosis assays or vaccines are fundamental. Therefore, this study aimed to use bioinformatics software to evaluate the critical chemical characteristics of unknown proteins of C. pseudotuberculosis by selecting them for heterologous expression in Escherichia coli. For this purpose, six protein sequences of ascorbate transporter subunit, UPF protein, MMPL family transporter, Ribonuclease, Iron ABC transporter domain-containing permease, and fimbrial subunit were obtained. In silico analyses were performed using amino acid sequences to access immunodominant epitopes and their antigenic and allergenic potential and physicochemical characterization. The expressed proteins were used as an antigen for serological diagnosis by ELISA. All proteins showed distinct immunodominant epitopes and potential antigenic characteristics. The only proteins expressed were PTS and Ribonuclease. In parallel, we expressed CP40 and all were used with ELISA antigen in 49 CLA positive sera and 26 CLA negative sera. The proteins alone showed 100% sensitivity and 96.2%, 92.3%, and 88.5% specificity for rPTS, rRibonuclease, and rCP40, respectively. When proteins were combined, they showed 100% sensitivity and 84.6%, 92.3%, 88.5%, and 92.3% specificity for rPTS/rCp40, rRibonuclease/rCP40, rPTS/rRibonuclease, and rPTS/rRibonuclease/rCP40, respectively. The results of this study show an excellent correlation of sensitivity and specificity with all proteins. None of the specificity values preclude the potential of rPTS, rRibonuclease, or rCP40 for use in ELISA diagnostic assays since the results of this work are superior to those of other studies on CLA diagnosis described in the literature. |
| format | Article |
| id | doaj-art-068992a80cab40b39e01ac41aa42c40d |
| institution | Kabale University |
| issn | 2090-8113 2042-0048 |
| language | English |
| publishDate | 2021-01-01 |
| publisher | Wiley |
| record_format | Article |
| series | Veterinary Medicine International |
| spelling | doaj-art-068992a80cab40b39e01ac41aa42c40d2025-02-03T01:08:48ZengWileyVeterinary Medicine International2090-81132042-00482021-01-01202110.1155/2021/99317319931731In Silico Screening of Putative Corynebacterium pseudotuberculosis Antigens and Serological Diagnosis for Caseous Lymphadenitis in Sheep by Enzyme-Linked Immunosorbent AssayDaniela Droppa-Almeida0Caroline de Santana Ferreira1Ioná Brito2Sibele Borsuk3Jorge Alberto López Rodríguez4Isabel Bezerra Lima-Verde5Francine Ferreira Padilha6Laboratório de Biomateriais, Instituto de Tecnologia e Pesquisa, Universidade Tiradentes, Aracaju, Sergipe, BrazilLaboratório de Biomateriais, Instituto de Tecnologia e Pesquisa, Universidade Tiradentes, Aracaju, Sergipe, BrazilLaboratório de Biomateriais, Instituto de Tecnologia e Pesquisa, Universidade Tiradentes, Aracaju, Sergipe, BrazilLaboratório de Biotecnologia Infecto-parasitária, Centro de Desenvolvimento Tecnológico, Universidade Federal de Pelotas, Pelotas, Rio Grande do Sul, BrazilLaboratório de Biologia Molecular, Instituto de Tecnologia e Pesquisa, Universidade Tiradentes, Aracaju, Sergipe, BrazilLaboratório de Biomateriais, Instituto de Tecnologia e Pesquisa, Universidade Tiradentes, Aracaju, Sergipe, BrazilLaboratório de Biomateriais, Instituto de Tecnologia e Pesquisa, Universidade Tiradentes, Aracaju, Sergipe, BrazilCorynebacterium pseudotuberculosis is the etiologic agent of Caseous Lymphadenitis (CLA), a disease leading to severe damage in sheep and goats farming due to the lack of serological diagnosis, treatment, and effective prophylaxis. In this context, several strategies in an attempt to discover new antigens to compose diagnosis assays or vaccines are fundamental. Therefore, this study aimed to use bioinformatics software to evaluate the critical chemical characteristics of unknown proteins of C. pseudotuberculosis by selecting them for heterologous expression in Escherichia coli. For this purpose, six protein sequences of ascorbate transporter subunit, UPF protein, MMPL family transporter, Ribonuclease, Iron ABC transporter domain-containing permease, and fimbrial subunit were obtained. In silico analyses were performed using amino acid sequences to access immunodominant epitopes and their antigenic and allergenic potential and physicochemical characterization. The expressed proteins were used as an antigen for serological diagnosis by ELISA. All proteins showed distinct immunodominant epitopes and potential antigenic characteristics. The only proteins expressed were PTS and Ribonuclease. In parallel, we expressed CP40 and all were used with ELISA antigen in 49 CLA positive sera and 26 CLA negative sera. The proteins alone showed 100% sensitivity and 96.2%, 92.3%, and 88.5% specificity for rPTS, rRibonuclease, and rCP40, respectively. When proteins were combined, they showed 100% sensitivity and 84.6%, 92.3%, 88.5%, and 92.3% specificity for rPTS/rCp40, rRibonuclease/rCP40, rPTS/rRibonuclease, and rPTS/rRibonuclease/rCP40, respectively. The results of this study show an excellent correlation of sensitivity and specificity with all proteins. None of the specificity values preclude the potential of rPTS, rRibonuclease, or rCP40 for use in ELISA diagnostic assays since the results of this work are superior to those of other studies on CLA diagnosis described in the literature.http://dx.doi.org/10.1155/2021/9931731 |
| spellingShingle | Daniela Droppa-Almeida Caroline de Santana Ferreira Ioná Brito Sibele Borsuk Jorge Alberto López Rodríguez Isabel Bezerra Lima-Verde Francine Ferreira Padilha In Silico Screening of Putative Corynebacterium pseudotuberculosis Antigens and Serological Diagnosis for Caseous Lymphadenitis in Sheep by Enzyme-Linked Immunosorbent Assay Veterinary Medicine International |
| title | In Silico Screening of Putative Corynebacterium pseudotuberculosis Antigens and Serological Diagnosis for Caseous Lymphadenitis in Sheep by Enzyme-Linked Immunosorbent Assay |
| title_full | In Silico Screening of Putative Corynebacterium pseudotuberculosis Antigens and Serological Diagnosis for Caseous Lymphadenitis in Sheep by Enzyme-Linked Immunosorbent Assay |
| title_fullStr | In Silico Screening of Putative Corynebacterium pseudotuberculosis Antigens and Serological Diagnosis for Caseous Lymphadenitis in Sheep by Enzyme-Linked Immunosorbent Assay |
| title_full_unstemmed | In Silico Screening of Putative Corynebacterium pseudotuberculosis Antigens and Serological Diagnosis for Caseous Lymphadenitis in Sheep by Enzyme-Linked Immunosorbent Assay |
| title_short | In Silico Screening of Putative Corynebacterium pseudotuberculosis Antigens and Serological Diagnosis for Caseous Lymphadenitis in Sheep by Enzyme-Linked Immunosorbent Assay |
| title_sort | in silico screening of putative corynebacterium pseudotuberculosis antigens and serological diagnosis for caseous lymphadenitis in sheep by enzyme linked immunosorbent assay |
| url | http://dx.doi.org/10.1155/2021/9931731 |
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