Development and Validation of a PCR-ELISA for the Diagnosis of Symptomatic and Asymptomatic Infection by Leishmania (Leishmania) infantum .

Autor: Medeiros FA; Centro de Pesquisas René Rachou, Oswaldo Cruz Foundation (Fiocruz), Belo Horizonte, MG, Brazil., Gomes LI; Centro de Pesquisas René Rachou, Oswaldo Cruz Foundation (Fiocruz), Belo Horizonte, MG, Brazil., Oliveira E; Centro de Pesquisas René Rachou, Oswaldo Cruz Foundation (Fiocruz), Belo Horizonte, MG, Brazil., de Souza CS; Centro de Pesquisas René Rachou, Oswaldo Cruz Foundation (Fiocruz), Belo Horizonte, MG, Brazil., Mourão MV; Centro de Pesquisas René Rachou, Oswaldo Cruz Foundation (Fiocruz), Belo Horizonte, MG, Brazil., Cota GF; Centro de Pesquisas René Rachou, Oswaldo Cruz Foundation (Fiocruz), Belo Horizonte, MG, Brazil., Marques LH; Departamento de Parasitologia, Instituto de Ciências Biológicas, Universidade Federal de Minas Gerais, Belo Horizonte, MG, Brazil., Carneiro M; Departamento de Parasitologia, Instituto de Ciências Biológicas, Universidade Federal de Minas Gerais, Belo Horizonte, MG, Brazil., Rabello A; Centro de Pesquisas René Rachou, Oswaldo Cruz Foundation (Fiocruz), Belo Horizonte, MG, Brazil.
Jazyk: angličtina
Zdroj: Journal of tropical medicine [J Trop Med] 2017; Vol. 2017, pp. 7364854. Date of Electronic Publication: 2017 Jan 09.
DOI: 10.1155/2017/7364854
Abstrakt: A kDNA PCR enzyme-linked immunosorbent assay ( kDNA PCR-ELISA) for the diagnosis of human visceral leishmaniasis (HVL) was developed. The detection limit of the reaction, precision measurements, and cut-off of the kDNA PCR-ELISA were defined in a proof-of-concept phase. A reference strain of Leishmania (Leishmania) infantum and 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 with Leishmania /HIV coinfection, 40 healthy individuals, and 33 asymptomatic individuals living in endemic areas. The kDNA PCR-ELISA exhibited satisfactory precision, with a detection limit of 0.07 fg of DNA from L . (L . ) infantum and 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%). The kDNA PCR-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.
Competing Interests: The authors declare that they have no competing interests.
Databáze: MEDLINE
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