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Biosens Bioelectron ; 86: 346-352, 2016 Dec 15.
Artigo em Inglês | MEDLINE | ID: mdl-27393827

RESUMO

Electrochemiluminescence (ECL) has been widely rendered for nucleic acid testing. Here, we integrate loop-mediated isothermal amplification (LAMP) with ECL technique for DNA detection and quantification. The target LAMP DNA bound electrostatically with [Ru(bpy)3](+2) on the carbon electrode surface, and an ECL reaction was triggered by tripropylamine (TPrA) to yield luminescence. We illustrated this method as a new and highly sensitive strategy for the detection of sequence-specific DNA from different meat species at picogram levels. The proposed strategy renders the signal amplification capacities of TPrA and combines LAMP with inherently high sensitivity of the ECL technique, to facilitate the detection of low quantities of DNA. By leveraging this technique, target DNA of Sus scrofa (pork) meat was detected as low as 1pg/µL (3.43×10(-1)copies/µL). In addition, the proposed technique was applied for detection of Bacillus subtilis DNA samples and detection limit of 10pg/µL (2.2×10(3)copies/µL) was achieved. The advantages of being isothermal, sensitive and robust with ability for multiplex detection of bio-analytes makes this method a facile and appealing sensing modality in hand-held devices to be used at the point-of-care (POC).


Assuntos
Condutometria/instrumentação , DNA/genética , Substâncias Luminescentes/química , Medições Luminescentes/instrumentação , Técnicas de Amplificação de Ácido Nucleico/instrumentação , Análise de Sequência de DNA/instrumentação , DNA/análise , Desenho de Equipamento , Análise de Falha de Equipamento , Ácidos Nucleicos , Reprodutibilidade dos Testes , Sensibilidade e Especificidade , Coloração e Rotulagem
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