title: Novel recombinant multiepitope proteins for the diagnosis of asymptomatic leishmania infantum-infected dogs.
abstract: BACKGROUND
  Visceral leishmaniasis is the most severe form of leishmaniasis. Worldwide, approximately 20% of zoonotic human visceral leishmaniasis is caused by Leishmania infantum, also known as Leishmania chagasi in Latin America. Current diagnostic methods are not accurate enough to identify Leishmania-infected animals and may compromise the effectiveness of disease control. Therefore, we aimed to produce and test two recombinant multiepitope proteins as a means to improve and increase accuracy in the diagnosis of canine visceral leishmaniasis (CVL).
  
  
  RESULTS
  Ten antigenic peptides were identified by CVL ELISA in previous work. In the current proposal, the coding sequences of these ten peptides were assembled into a synthetic gene. Furthermore, other twenty peptides were selected from work by our group where good B and T cell epitopes were mapped. The coding sequences of these peptides were also assembled into a synthetic gene. Both genes have been cloned and expressed in Escherichia coli, producing two multiepitope recombinant proteins, PQ10 and PQ20. These antigens have been used in CVL ELISA and were able to identify asymptomatic dogs (80%) more effectively than EIE-LVC kit, produced by Bio-Manguinhos (0%) and DPP kit (10%). Moreover, our recombinant proteins presented an early detection (before PCR) of infected dogs, with positivities ranging from 23% to 65%, depending on the phase of infection in which sera were acquired.
  
  
  CONCLUSIONS
  Our study shows that ELISA using the multiepitope proteins PQ10 and PQ20 has great potential in early CVL diagnosis. The use of these proteins in other methodologies, such as immunochromatographic tests, could be beneficial mainly for the detection of asymptomatic dogs.
authors.affiliation: Universidade Federal de Minas Gerais, Instituto de Ciências Biológicas, Departamento de Parasitologia, Belo Horizonte, Minas Gerais, Brasil.
authors.forename: Angélica Rosa
authors.initials: AR
authors.lastname: Faria
authors.affiliation: Universidade Federal de Minas Gerais, Instituto de Ciências Biológicas, Departamento de Parasitologia, Belo Horizonte, Minas Gerais, Brasil.
authors.forename: Luciano
authors.initials: L
authors.lastname: de Castro Veloso
authors.affiliation: Universidade Federal de Ouro Preto, Núcleo de Pesquisa em Ciências Biológicas, Ouro Preto, Minas Gerais, Brasil.
authors.forename: Wendel
authors.initials: W
authors.lastname: Coura-Vital
authors.affiliation: Universidade Federal de Ouro Preto, Núcleo de Pesquisa em Ciências Biológicas, Ouro Preto, Minas Gerais, Brasil.
authors.forename: Alexandre Barbosa
authors.initials: AB
authors.lastname: Reis
authors.affiliation: Universidade Federal de Minas Gerais, Instituto de Ciências Biológicas, Departamento de Bioquímica e Imunologia, Belo Horizonte, Minas Gerais, Brasil.
authors.forename: Leonardo Miranda
authors.initials: LM
authors.lastname: Damasceno
authors.affiliation: Universidade Federal de Minas Gerais, Instituto de Ciências Biológicas, Departamento de Bioquímica e Imunologia, Belo Horizonte, Minas Gerais, Brasil; Centro de Pesquisas René Rachou - Fundação Oswaldo Cruz, Belo Horizonte, Minas Gerais, Brasil; University of Massachusetts Medical School, Division of Infectious Diseases and Immunology, Worcester, Massachusetts, United States of America.
authors.forename: Ricardo T
authors.initials: RT
authors.lastname: Gazzinelli
authors.affiliation: Universidade Federal de Minas Gerais, Instituto de Ciências Biológicas, Departamento de Parasitologia, Belo Horizonte, Minas Gerais, Brasil.
authors.forename: Hélida M
authors.initials: HM
authors.lastname: Andrade
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000953:Antigens, Protozoan; D004283:Dog Diseases / Q000175:diagnosis / Q000469:parasitology*; D004285:Dogs; D004797:Enzyme-Linked Immunosorbent Assay / Q000662:veterinary; D000939:Epitopes; D018314:Leishmania infantum; D007898:Leishmaniasis, Visceral / Q000097:blood / Q000175:diagnosis / Q000469:parasitology / Q000662:veterinary*; D011994:Recombinant Proteins / Q000378:metabolism*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity
chemical_list: D000953:Antigens, Protozoan; D000939:Epitopes; D011994:Recombinant Proteins