title: Use of Recombinant Antigens for Sensitive Serodiagnosis of American Tegumentary Leishmaniasis Caused by Different Leishmania Species.
abstract: American tegumentary leishmaniasis (ATL) (also known as cutaneous leishmaniasis [CL]) is caused by various species of protozoa of the genus Leishmania The diagnosis is achieved on a clinical, epidemiological, and pathological basis, supported by positive parasitological exams and demonstration of leishmanin delayed-type hypersensitivity. Serological assays are not routinely used in the diagnosis because many are considered to have low sensitivity and the particular Leishmania species causing the disease can lead to variable performance. In the present study, we generated recombinant versions of two highly conserved Leishmania proteins, Leishmania (Viannia) braziliensis-derived Lb8E and Lb6H, and evaluated both in enzyme-linked immunosorbent assays (ELISA). Recombinant Lb6H (rLb6H) had better performance and reacted with 100.0% of the ATL and 89.4% of the VL samples. These reactions with rLb6H were highly specific (98.5%) when compared against those for samples from healthy control individuals. We then assessed rLb6H against sera from ATL patients infected with different species of Leishmania prevalent in Brazil [Leishmania (Leishmania) amazonensis, L (Viannia) braziliensis, and L (V) guyanensis] and samples from patients with other infectious diseases. In analyses of 500 sera, ELISA using rLb6H detected all 219 ATL samples (sensitivity of 100.0%) with an overall specificity of 93.9% (considering healthy individuals and other infectious diseases patients). Only a minority of samples from Chagas disease patients possessed antibodies against rLb6H, and all of these responses were low (with a highest reactivity index of 2.2). Taken together, our data support further evaluation of rLb6H and the potential for its routine use in the serological diagnosis of ATL.
authors.affiliation: Instituto de Medicina Tropical de São Paulo, Universidade de São Paulo, São Paulo, Brazil.
authors.forename: Camila Massae
authors.initials: CM
authors.lastname: Sato
authors.affiliation: Instituto de Medicina Tropical de São Paulo, Universidade de São Paulo, São Paulo, Brazil.
authors.forename: Maria Carmen Arroyo
authors.initials: MC
authors.lastname: Sanchez
authors.affiliation: Instituto de Medicina Tropical de São Paulo, Universidade de São Paulo, São Paulo, Brazil.
authors.forename: Beatriz Julieta
authors.initials: BJ
authors.lastname: Celeste
authors.affiliation: Infectious Disease Research Institute, Seattle, Washington, USA.
authors.forename: Malcolm S
authors.initials: MS
authors.lastname: Duthie
authors.affiliation: Infectious Disease Research Institute, Seattle, Washington, USA.
authors.forename: Jeffrey
authors.initials: J
authors.lastname: Guderian
authors.affiliation: Infectious Disease Research Institute, Seattle, Washington, USA.
authors.forename: Steven G
authors.initials: SG
authors.lastname: Reed
authors.affiliation: Department of Immunology, Aggeu Magalhães Research Center-Oswaldo Cruz Foundation (FIOCRUZ), Recife, Pernambuco, Brazil.
authors.forename: Maria Edileuza Felinto
authors.initials: ME
authors.lastname: de Brito
authors.affiliation: Instituto Evandro Chagas, Belém, Pará, Brazil.
authors.forename: Marliane Batista
authors.initials: MB
authors.lastname: Campos
authors.affiliation: Fundação de Medicina Tropical Dr. Heitor Vieira Dourado, Manaus, Amazonas, Brazil.
authors.forename: Helia Valeria
authors.initials: HV
authors.lastname: de Souza Encarnação
authors.affiliation: Fundação de Medicina Tropical Dr. Heitor Vieira Dourado, Manaus, Amazonas, Brazil.|Universidade do Estado do Amazonas, Amazonas, Brazil.
authors.forename: Jorge
authors.initials: J
authors.lastname: Guerra
authors.affiliation: Fundação de Medicina Tropical Dr. Heitor Vieira Dourado, Manaus, Amazonas, Brazil.
authors.forename: Tirza Gabrielle Ramos
authors.initials: TG
authors.lastname: de Mesquita
authors.affiliation: Fundação de Medicina Tropical Dr. Heitor Vieira Dourado, Manaus, Amazonas, Brazil.
authors.forename: Suzana Kanawati
authors.initials: SK
authors.lastname: Pinheiro
authors.affiliation: Fundação de Medicina Tropical Dr. Heitor Vieira Dourado, Manaus, Amazonas, Brazil.|Universidade Nilton Lins, Manaus, Amazonas, Brazil.
authors.forename: Rajendranath
authors.initials: R
authors.lastname: Ramasawmy
authors.affiliation: Instituto Evandro Chagas, Belém, Pará, Brazil.|Universidade Federal do Pará/Núcleo de Medicina Tropical, UFPA/NMT, Pará, Brazil.
authors.forename: Fernando Tobias
authors.initials: FT
authors.lastname: Silveira
authors.affiliation: Instituto de Medicina Tropical de São Paulo, Universidade de São Paulo, São Paulo, Brazil.|Department of Immunology, Aggeu Magalhães Research Center-Oswaldo Cruz Foundation (FIOCRUZ), Recife, Pernambuco, Brazil.
authors.forename: Marina
authors.initials: M
authors.lastname: de Assis Souza
authors.affiliation: Instituto de Medicina Tropical de São Paulo, Universidade de São Paulo, São Paulo, Brazil hgoto@usp.br.|Faculdade de Medicina da Universidade de São Paulo, São Paulo, Brazil.
authors.forename: Hiro
authors.identifier: 0000-0001-7048-562X
authors.initials: H
authors.lastname: Goto
mesh_terms: D000293:Adolescent; D000328:Adult; D000368:Aged; D000913:Antibodies, Protozoan / Q000097:blood*; D000953:Antigens, Protozoan / Q000235:genetics / Q000276:immunology*; D002648:Child; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D005260:Female; D006801:Humans; D007891:Leishmania / Q000276:immunology*; D016773:Leishmaniasis, Cutaneous / Q000175:diagnosis*; D008297:Male; D008875:Middle Aged; D011994:Recombinant Proteins / Q000235:genetics / Q000276:immunology*; D012680:Sensitivity and Specificity; D012698:Serologic Tests / Q000379:methods*; D055815:Young Adult
chemical_list: D000913:Antibodies, Protozoan; D000953:Antigens, Protozoan; D011994:Recombinant Proteins
keywords: ELISA; diagnostics; immunoserology; recombinant antigen; tegumentary leishmaniasis