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 Jul:147:102505. doi: 10.1016/j.tube.2024.102505. Epub 2024 Mar 30.

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.

MeSH terms

  • Antibodies, Bacterial / blood
  • Antigens, Bacterial* / genetics
  • Antigens, Bacterial* / immunology
  • Bacterial Proteins* / genetics
  • Bacterial Proteins* / immunology
  • Enzyme-Linked Immunosorbent Assay* / methods
  • Epitopes, B-Lymphocyte* / immunology
  • Female
  • Humans
  • Leprosy, Multibacillary* / diagnosis
  • Leprosy, Multibacillary* / immunology
  • Leprosy, Paucibacillary* / diagnosis
  • Leprosy, Paucibacillary* / immunology
  • Male
  • Mycobacterium leprae* / genetics
  • Mycobacterium leprae* / immunology
  • Predictive Value of Tests
  • Recombinant Fusion Proteins / immunology
  • Recombinant Proteins / genetics
  • Recombinant Proteins / immunology
  • Sensitivity and Specificity
  • Serologic Tests* / methods

Substances

  • Epitopes, B-Lymphocyte
  • Antigens, Bacterial
  • Bacterial Proteins
  • Antibodies, Bacterial
  • Recombinant Fusion Proteins
  • Recombinant Proteins