Development and Validation of a PCR-ELISA for the Diagnosis of Symptomatic and Asymptomatic Infection byLeishmania (Leishmania) infantum.
AkDNAPCR enzyme-linked immunosorbent assay (kDNAPCR-ELISA) for the diagnosis of human visceral leishmaniasis (HVL) was developed. The detection limit of the reaction, precision measurements, and cut-off of thekDNAPCR-ELISA were defined in a proof-of-concept phase. A reference strain ofLeishmania (Leishmania) infantumand a bank of 14 peripheral blood samples from immunocompetent patients with VL were characterized using techniques considered gold standards, and 11 blood samples obtained from healthy individuals of an endemic area were also assessed. Phase II evaluation determined the performance of the assay in peripheral blood samples from 105 patients with VL (adults and children), 25 patients withLeishmania/HIV coinfection, 40 healthy individuals, and 33 asymptomatic individuals living in endemic areas. ThekDNAPCR-ELISA exhibited satisfactory precision, with a detection limit of 0.07 fg of DNA fromL.(L.) infantumand 1 parasite/mL blood. The overall sensitivity of the assay for all groups studied was 100% (95% confidence interval [CI]: 97.1-100%), and the specificity was 95% (95% CI: 83.5-98.6%). ThekDNAPCR-ELISA was shown to be a useful tool for VL symptomatic and asymptomatic individuals diagnosis and its use in endemic countries may help monitor control interventions.
Centro de Pesquisas René Rachou, Oswaldo Cruz Foundation (Fiocruz), Belo Horizonte, MG, Brazil.
Fernanda Alvarenga Cardoso
FA
Medeiros
Centro de Pesquisas René Rachou, Oswaldo Cruz Foundation (Fiocruz), Belo Horizonte, MG, Brazil.
Luciana Inácia
LI
Gomes
Centro de Pesquisas René Rachou, Oswaldo Cruz Foundation (Fiocruz), Belo Horizonte, MG, Brazil.
Edward
0000-0002-0042-5178
E
Oliveira
Centro de Pesquisas René Rachou, Oswaldo Cruz Foundation (Fiocruz), Belo Horizonte, MG, Brazil.
Carolina Senra Alves
CS
de Souza
Centro de Pesquisas René Rachou, Oswaldo Cruz Foundation (Fiocruz), Belo Horizonte, MG, Brazil.
Maria Vitória
MV
Mourão
Centro de Pesquisas René Rachou, Oswaldo Cruz Foundation (Fiocruz), Belo Horizonte, MG, Brazil.
Gláucia Fernandes
GF
Cota
Departamento de Parasitologia, Instituto de Ciências Biológicas, Universidade Federal de Minas Gerais, Belo Horizonte, MG, Brazil.
Letícia Helena Dos Santos
LH
Marques
Departamento de Parasitologia, Instituto de Ciências Biológicas, Universidade Federal de Minas Gerais, Belo Horizonte, MG, Brazil.
Mariângela
M
Carneiro
Centro de Pesquisas René Rachou, Oswaldo Cruz Foundation (Fiocruz), Belo Horizonte, MG, Brazil.
Ana
0000-0001-5821-4069
A
Rabello