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Anal Chim Acta ; 1041: 40-49, 2018 Dec 24.
Artigo em Inglês | MEDLINE | ID: mdl-30340689

RESUMO

Detecting and monitoring the pathogens with high selectivity and sensitivity is critical for public health. In the present study, we demonstrated a specific analytical strategy for sensitive detection of Leishmania infantum genome. The developed sensor utilized toluidine blue as a hybridization indicator and a Leishmania infantum-specific capture DNA sequence immobilized on a high-surface area gold nanostructure as an electrochemical transducer. The produced analytical response was based on the hybridization of the single-stranded DNA from the target with the immobilized DNA sequence at the electrode surface. The developed DNA sensor in this study was successfully employed to detect a synthetic Leishmania infantum target sequence in a wide concentration range from 1 × 10-18 to 1 × 10-10 mol L-1 with a detection limit of 0.2 amol L-1 with the ability to discriminate the target sequence from mismatched sequences. Moreover, the designed DNA sensor showed a good reproducibility and stability during repeated regeneration and hybridization cycles. The DNA sensor could detect Leishmania infantum genome in a wide concentration range from 15 to 50 ng µL-1 with a detection limit of 29 ng µL-1. Furthermore, clinical trials confirmed the applicability of the developed DNA sensor for practical applications.


Assuntos
DNA Bacteriano/análise , Técnicas Eletroquímicas , Ouro/química , Leishmania infantum/isolamento & purificação , Nanopartículas Metálicas/química , DNA Bacteriano/metabolismo , DNA de Cadeia Simples/química , DNA de Cadeia Simples/metabolismo , Eletrodos , Genoma Bacteriano , Ácidos Nucleicos Imobilizados/química , Ácidos Nucleicos Imobilizados/metabolismo , Leishmania infantum/genética , Limite de Detecção , Hibridização de Ácido Nucleico , Reprodutibilidade dos Testes
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