title: Phage display and synthetic peptides as promising biotechnological tools for the serological diagnosis of leprosy.
abstract: BACKGROUND
  The diagnosis of leprosy is primarily based on clinical manifestations, and there is no widely available laboratory test for the early detection of this disease, which is caused by Mycobacterium leprae. In fact, early detection and treatment are the key elements to the successful control of leprosy.
  
  
  RESULTS
  Peptide ligands for antibodies from leprosy patients were selected from phage-displayed peptide libraries. Three peptide sequences expressed by reactive phage clones were chemically synthesized. Serological assays that used synthetic peptides were evaluated using serum samples from leprosy patients, household contacts (HC) of leprosy patients, tuberculosis patients and endemic controls (EC). A pool of three peptides identified 73.9% (17/23) of multibacillary (MB) leprosy patients using an enzyme-linked immunosorbent assay (ELISA). These peptides also showed some seroreactivities to the HC and EC individuals. The peptides were not reactive to rabbit polyclonal antisera against the different environmental mycobacteria. The same peptides that were conjugated to the carrier protein bovine serum albumin (BSA) induced the production of antibodies in the mice. The anti-peptide antibodies that were used in the Western blotting analysis of M. leprae crude extracts revealed a single band of approximately 30 kDa in one-dimensional electrophoresis and four 30 kDa isoforms in the two-dimensional gel. The Western blotting data indicated that the three peptides are derived from the same bacterial protein.
  
  
  CONCLUSIONS
  These new antigens may be useful in the diagnosis of MB leprosy patients. Their potentials as diagnostic reagents must be more extensively evaluated in future studies using a large panel of positive and negative sera. Furthermore, other test approaches using peptides should be assessed to increase their sensitivity and specificity in detecting leprosy patients. We have revealed evidence in support of phage-displayed peptides as promising biotechnological tools for the design of leprosy diagnostic serological assays.
authors.affiliation: Department of Biotechnology and Bioprocess Engineering, Federal University of Parana, Curitiba, Parana, Brazil.
authors.forename: Silvana Maria
authors.initials: SM
authors.lastname: Alban
authors.affiliation: Basic Pathology Department, Federal University of Parana, Curitiba, Parana, Brazil.
authors.forename: Juliana Ferreira
authors.initials: JF
authors.lastname: de Moura
authors.affiliation: Department of Biotechnology and Bioprocess Engineering, Federal University of Parana, Curitiba, Parana, Brazil.
authors.forename: Vanete
authors.initials: V
authors.lastname: Thomaz-Soccol
authors.affiliation: Immunology Department, Tropical Pathology and Public Health Institute, Federal University of Goias, Goiania, Goias, Brazil.
authors.forename: Samira
authors.initials: S
authors.lastname: Bührer Sékula
authors.affiliation: Immunology Department, Tropical Pathology and Public Health Institute, Federal University of Goias, Goiania, Goias, Brazil.
authors.forename: Larissa Magalhães
authors.initials: LM
authors.lastname: Alvarenga
authors.affiliation: Health Sciences Postgraduate Program, School of Medicine, Pontifical Catholic University of Parana, Curitiba, Parana, Brazil.
authors.forename: Marcelo Távora
authors.initials: MT
authors.lastname: Mira
authors.affiliation: Department of Biochemistry and Immunology, Federal University of Minas Gerais, Belo Horizonte, Minas Gerais, Brazil.
authors.forename: Carlos Chávez
authors.initials: CC
authors.lastname: Olortegui
authors.affiliation: Center for Production and Research of Immunobiological Products, Parana State Department of Health, Piraquara, Parana, Brazil.
authors.forename: João Carlos
authors.initials: JC
authors.lastname: Minozzo
mesh_terms: D000818:Animals; D000907:Antibodies, Bacterial / Q000097:blood* / Q000737:chemistry; D002417:Cattle; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D007918:Leprosy / Q000097:blood* / Q000175:diagnosis*; D051379:Mice; D009166:Mycobacterium leprae; D019151:Peptide Library; D011817:Rabbits
chemical_list: D000907:Antibodies, Bacterial; D019151:Peptide Library