title: New serological tools for improved diagnosis of human tegumentary leishmaniasis.
abstract: Human tegumentary leishmaniasis (HTL), characterized by skin ulcers that may spread and cause dreadful and massive tissue destruction of the nose and mouth, is considered a neglected tropical disease, and it is a serious threat to global health due to its continuous expansion, favored by the lifecycle of its causative organism that is maintained in domestic animal reservoirs and anthropophilic sand fly species. Serodiagnosis of HTL is a great challenge due to many biological factors, including hampered specificity and/or sensitivity. This investigation addresses the unmet need for new diagnostic markers of HTL, and describes a simple platform to improve the serodiagnosis. A constrained conformational phage display random peptide library combined with a magnetic microsphere-based subtraction strategy was used to identify ligands with potential diagnostic applications. Six clones were selected against IgG antibodies from HTL patients, characterized by sequencing and confirmed by a phage-ELISA using sera from patients developing visceral leishmaniasis (n=20), Chagas disease (n=10), mucosal (n=30) and cutaneous (n=20) leishmaniasis; as well as from healthy subjects living in endemic (n=20) and non-endemic (n=30) areas of leishmaniasis. A wild-type M13-phage clone and a soluble Leishmania antigenic extract were used as negative and positive controls, respectively. Three clones reached 100% sensitivity and specificity, without any cross-reactivity with sera from patients with leishmaniasis-related diseases. Briefly, we describe for the first time a set of serological markers based on three immunodominant mimotopes that showed 100% accuracy, and that could be used in a phage-ELISA assay for the HTL serodiagnosis.
authors.affiliation: Programa de Pós-Graduação em Ciências Saúde: Infectologia e Medicina Tropical, Faculdade de Medicina, Universidade Federal de Minas Gerais, Av. Antônio Carlos, 6627, 30130-100 Belo Horizonte, Minas Gerais, Brazil.
authors.forename: Lourena E
authors.initials: LE
authors.lastname: Costa
authors.affiliation: Programa de Pós-Graduação em Ciências Saúde: Infectologia e Medicina Tropical, Faculdade de Medicina, Universidade Federal de Minas Gerais, Av. Antônio Carlos, 6627, 30130-100 Belo Horizonte, Minas Gerais, Brazil.
authors.forename: Beatriz C S
authors.initials: BC
authors.lastname: Salles
authors.affiliation: Laboratório de Nanobiotecnologia, Instituto de Genética e Bioquímica, Universidade Federal de Uberlândia, Av. Amazonas s/n, Campus Umuarama, Bloco 2E, Sala 248, 38400-902 Uberlândia, Minas Gerais, Brazil.
authors.forename: Patrícia T
authors.initials: PT
authors.lastname: Alves
authors.affiliation: Laboratório de Nanobiotecnologia, Instituto de Genética e Bioquímica, Universidade Federal de Uberlândia, Av. Amazonas s/n, Campus Umuarama, Bloco 2E, Sala 248, 38400-902 Uberlândia, Minas Gerais, Brazil.
authors.forename: Ana C S
authors.initials: AC
authors.lastname: Dias
authors.affiliation: Laboratório de Nanobiotecnologia, Instituto de Genética e Bioquímica, Universidade Federal de Uberlândia, Av. Amazonas s/n, Campus Umuarama, Bloco 2E, Sala 248, 38400-902 Uberlândia, Minas Gerais, Brazil.
authors.forename: Emília R
authors.initials: ER
authors.lastname: Vaz
authors.affiliation: Departamento de Patologia Clínica, Coltec, Universidade Federal de Minas Gerais, Av. Antônio Carlos, 6627, 31270-901 Belo Horizonte, Minas Gerais, Brazil.
authors.forename: Fernanda F
authors.initials: FF
authors.lastname: Ramos
authors.affiliation: Departamento de Patologia Clínica, Coltec, Universidade Federal de Minas Gerais, Av. Antônio Carlos, 6627, 31270-901 Belo Horizonte, Minas Gerais, Brazil.
authors.forename: Daniel
authors.initials: D
authors.lastname: Menezes-Souza
authors.affiliation: Departamento de Patologia Clínica, Coltec, Universidade Federal de Minas Gerais, Av. Antônio Carlos, 6627, 31270-901 Belo Horizonte, Minas Gerais, Brazil.
authors.forename: Mariana C
authors.initials: MC
authors.lastname: Duarte
authors.affiliation: Departamento de Patologia Clínica, Coltec, Universidade Federal de Minas Gerais, Av. Antônio Carlos, 6627, 31270-901 Belo Horizonte, Minas Gerais, Brazil.
authors.forename: Bruno M
authors.initials: BM
authors.lastname: Roatt
authors.affiliation: Programa de Pós-Graduação em Ciências Saúde: Infectologia e Medicina Tropical, Faculdade de Medicina, Universidade Federal de Minas Gerais, Av. Antônio Carlos, 6627, 30130-100 Belo Horizonte, Minas Gerais, Brazil.
authors.forename: Miguel A
authors.initials: MA
authors.lastname: Chávez-Fumagalli
authors.affiliation: Departamento de Bioquímica e Imunologia, Instituto de Ciências Biológicas, Universidade Federal de Minas Gerais, Av. Antônio Carlos, 6627, 31270-901 Belo Horizonte, Minas Gerais, Brazil.
authors.forename: Carlos A P
authors.initials: CA
authors.lastname: Tavares
authors.affiliation: Programa de Pós-Graduação em Ciências Saúde: Infectologia e Medicina Tropical, Faculdade de Medicina, Universidade Federal de Minas Gerais, Av. Antônio Carlos, 6627, 30130-100 Belo Horizonte, Minas Gerais, Brazil.
authors.forename: Denise U
authors.initials: DU
authors.lastname: Gonçalves
authors.affiliation: Programa de Pós-Graduação em Ciências Saúde: Infectologia e Medicina Tropical, Faculdade de Medicina, Universidade Federal de Minas Gerais, Av. Antônio Carlos, 6627, 30130-100 Belo Horizonte, Minas Gerais, Brazil.
authors.forename: Manoel O C
authors.initials: MO
authors.lastname: Rocha
authors.affiliation: Laboratório de Nanobiotecnologia, Instituto de Genética e Bioquímica, Universidade Federal de Uberlândia, Av. Amazonas s/n, Campus Umuarama, Bloco 2E, Sala 248, 38400-902 Uberlândia, Minas Gerais, Brazil; Department of Medical Microbiology and Immunology, University of California-Davis, 95616 Davis, CA, USA. Electronic address: lrgoulart@ufu.br.
authors.forename: Luiz Ricardo
authors.initials: LR
authors.lastname: Goulart
authors.affiliation: Programa de Pós-Graduação em Ciências Saúde: Infectologia e Medicina Tropical, Faculdade de Medicina, Universidade Federal de Minas Gerais, Av. Antônio Carlos, 6627, 30130-100 Belo Horizonte, Minas Gerais, Brazil. Electronic address: eduardoferrazcoelho@yahoo.com.br.
authors.forename: Eduardo A F
authors.initials: EA
authors.lastname: Coelho
mesh_terms: D000328:Adult; D000818:Animals; D000913:Antibodies, Protozoan / Q000097:blood; D000953:Antigens, Protozoan / Q000276:immunology; D001938:Brazil; D014355:Chagas Disease / Q000175:diagnosis; D003429:Cross Reactions; D004285:Dogs; D005260:Female; D006801:Humans; D007892:Leishmania braziliensis; D016773:Leishmaniasis, Cutaneous / Q000175:diagnosis*; D007898:Leishmaniasis, Visceral / Q000175:diagnosis; D008297:Male; D008875:Middle Aged; D012372:ROC Curve; D012680:Sensitivity and Specificity; D012698:Serologic Tests / Q000379:methods*; D055815:Young Adult
chemical_list: D000913:Antibodies, Protozoan; D000953:Antigens, Protozoan
keywords: Human tegumentary leishmaniasis; Mimotopes; Phage display; Phage-ELISA; Serodiagnosis; Specificity