Serodiagnosis of paucibacillary and multibacillary leprosy using a recombinant chimeric protein composed of specific B-cell epitopes derived from Mycobacterium leprae proteins

Bárbara P N Assis; Ana T Chaves; Daniela P Lage; Mariana M Cardoso; Camila S Freitas; Isabela A G Pereira; Raquel S B Câmara; Vívian T Martins; Ana Laura G de Oliveira; Ricardo A Machado-de-Ávila; Alexsandro S Galdino; Miguel A Chávez-Fumagalli; Myron Christodoulides; Denise U Gonçalves; Lílian L Bueno; Ricardo T Fujiwara; Eduardo A F Coelho; Manoel O da Costa Rocha

doi:10.1016/j.tube.2024.102505

Serodiagnosis of paucibacillary and multibacillary leprosy using a recombinant chimeric protein composed of specific B-cell epitopes derived from Mycobacterium leprae proteins

Tuberculosis (Edinb). 2024 Mar 30:147:102505. doi: 10.1016/j.tube.2024.102505. Online ahead of print.

Authors

Bárbara P N Assis¹, Ana T Chaves², Daniela P Lage², Mariana M Cardoso², Camila S Freitas², Isabela A G Pereira², Raquel S B Câmara², Vívian T Martins², Ana Laura G de Oliveira², Ricardo A Machado-de-Ávila³, Alexsandro S Galdino⁴, Miguel A Chávez-Fumagalli⁵, Myron Christodoulides⁶, Denise U Gonçalves², Lílian L Bueno⁷, Ricardo T Fujiwara⁷, Eduardo A F Coelho⁸, Manoel O da Costa Rocha²

Affiliations

¹ Programa de Pós-Graduação em Ciências da Saúde: Infectologia e Medicina Tropical, Faculdade de Medicina, Universidade Federal de Minas Gerais, Belo Horizonte, 30130-100, Minas Gerais, Brazil; Fundação Hospitalar do Estado de Minas Gerais, Hospital Eduardo de Menezes, Belo Horizonte, 30622-020, Minas Gerais, Brazil.
² Programa de Pós-Graduação em Ciências da Saúde: Infectologia e Medicina Tropical, Faculdade de Medicina, Universidade Federal de Minas Gerais, Belo Horizonte, 30130-100, Minas Gerais, Brazil.
³ Programa de Pós-Graduação em Ciências da Saúde, Universidade do Extremo Sul Catarinense, Criciúma, 88806-000, Santa Catarina, Brazil.
⁴ Laboratório de Biotecnologia de Microrganismos, Universidade Federal de São João Del-Rei, Divinópolis, 35.501-296, Minas Gerais e Instituto Nacional de Ciência e Tecnologia em Biotecnologia Industrial (INCT-BI), Brasil.
⁵ Computational Biology and Chemistry Research Group, Vicerrectorado de Investigación, Universidad Católica de Santa María, Urb. San José S/N, Umacollo, Arequipa, 04000, Peru.
⁶ Neisseria Research Group, Molecular Microbiology, School of Clinical and Experimental Sciences, University of Southampton Faculty of Medicine, Southampton General Hospital, Southampton, England, UK.
⁷ Programa de Pós-Graduação em Ciências da Saúde: Infectologia e Medicina Tropical, Faculdade de Medicina, Universidade Federal de Minas Gerais, Belo Horizonte, 30130-100, Minas Gerais, Brazil; Departamento de Parasitologia, Instituto de Ciências Biológicas, Universidade Federal de Minas Gerais, Belo Horizonte, 31270-901, Minas Gerais, Brazil.
⁸ Programa de Pós-Graduação em Ciências da Saúde: Infectologia e Medicina Tropical, Faculdade de Medicina, Universidade Federal de Minas Gerais, Belo Horizonte, 30130-100, Minas Gerais, Brazil; Departamento de Patologia Clínica, COLTEC, Universidade Federal de Minas Gerais, Av. Antônio Carlos, 6627, Belo Horizonte, Minas Gerais, Brazil. Electronic address: eduardoferrazcoelho@yahoo.com.br.

PMID: 38583359
DOI: 10.1016/j.tube.2024.102505

Abstract

Leprosy diagnosis is difficult due to the clinical similarity with other infectious diseases, and laboratory tests presents problems related to sensitivity and/or specificity. In this study, we used bioinformatics to assess Mycobacterium leprae proteins and formulated a chimeric protein that was tested as a diagnostic marker for the disease. The amino acid sequences from ML0008, ML0126, ML0308, ML1057, ML2028, ML2038, ML2498 proteins were evaluated, and the B-cell epitopes QASVAYPATSYADFRAHNHWWNGP, SLQRSISPNSYNTARVDP and QLLGQTADVAGAAKSGPVQPMGDRGSVSPVGQ were considered M. leprae-specific and used to construct the gene encoding the recombinant antigen. The gene was constructed, the recombinant protein was expressed, purified and tested in ELISA using 252 sera, which contained samples from multibacillary (MB) or paucibacillary (PB) leprosy patients, from their household contacts and healthy individuals, as well as from patients with Chagas disease, visceral and tegumentary leishmaniases (VL/TL), malaria, tuberculosis, and HIV. Sensitivity (Se) and specificity (Sp) for MB and PB samples compared to sera from both healthy subjects and individuals with cross-reactive diseases were 100%. The Se value for MB and PB samples compared to sera from household contacts was 100%, but Sp was 64%. In conclusion, data suggest that this protein could be considered in future studies for leprosy diagnosis.

Keywords: B-cell epitopes; Chimeric protein; Diagnosis; ELISA; Genomics; Leprosy.