期刊
SENSORS AND ACTUATORS B-CHEMICAL
卷 268, 期 -, 页码 438-445出版社
ELSEVIER SCIENCE SA
DOI: 10.1016/j.snb.2018.04.133
关键词
Electrochemical biosensor; Helicase; Isothermal amplification; Salmonella detection
资金
- Spanish Ministerio de Economiia y Competitividad [CTQ2015-63567-R]
- Principado de Asturias government [FC15-GRUPIN14-025]
- FEDER funds
Real-time PCR is the gold standard technique for the unambiguous detection of sequence-specific fragments of pathogen DNA in many fields but it does not always fulfill the extremely low detectability legislation demands. Herein we demonstrate that rationally designed genosensors can rival or even supersede it when combined with previous amplification strategies even isothermal ones. Among the latter, helicase-dependent amplification (HDA) is a simple and powerful DNA amplification using helicase instead of temperature cycles to unwind DNA; it follows an amplification scheme very similar to PCR, which could facilitate its widespread adoption for point-of-need applications. Unfortunately, the primer design for HDA is more restrictive than for PCR. We show that it is possible to adapt PCR primers to isothermal amplification of a DNA sequence specific for Salmonella. We design an electrochemical DNA sensor, based on a sandwich hybridization assay on the surface of indium-tin oxide electrodes, which allows monitoring both PCR and HDA amplicons. The resulting sequence-specific detection strategy not only increased the selectivity of the detection but also its sensitivity, improving real-time PCR in twofold and a hundred-fold when combined with HDA and PCR, respectively, achieving the single molecule detection. (c) 2018 Elsevier B.V. All rights reserved.
作者
我是这篇论文的作者
点击您的名字以认领此论文并将其添加到您的个人资料中。
推荐
暂无数据