Spehar-Délèze Anna-Maria, Gransee Rainer, Martinez-Montequin Sergio, Bejarano-Nosas Diego, Dulay Samuel, Julich Sandra, Tomaso Herbert, O'Sullivan Ciara K
Nanobiotechnology and Bioanalysis Group, INTERFIBIO Consolidated Research Group, Departament d'Enginyeria Quimica, Universitat Rovira i Virgili, Avinguda Països Catalans 26, 43007, Tarragona, Spain,
Anal Bioanal Chem. 2015 Sep;407(22):6657-67. doi: 10.1007/s00216-015-8831-y. Epub 2015 Jun 23.
Development of a fully automated electrochemiluminescence (ECL) DNA assay for multiplex detection of six biowarfare agents is described. Aminated-DNA capture probes were covalently immobilised on activated-carbon electrodes and subsequently hybridised to target strands. Detection was achieved via a sandwich-type assay after Ru(bpy)3(2+)-labelled reporter probes were hybridised to the formed probe-target complexes. The assay was performed in an automated microsystem in a custom designed ECL detection box with integrated fluidics, electronics,and movable photomultiplier detector. The obtained limits of detection were 0.6-1.2 nmol L(-1) for six targets ranging from 50 to 122 base pairs in size, with linear range 1-15 nmol L(-1). Non-specific adsorption and cross-reactivity were very low. Detection of six targets on a single chip was achieved with subnanomolar detection limits.
本文描述了一种用于多重检测六种生物战剂的全自动电化学发光(ECL)DNA检测方法。胺化DNA捕获探针共价固定在活性炭电极上,随后与靶链杂交。在钌(联吡啶)3(2+)标记的报告探针与形成的探针-靶标复合物杂交后,通过夹心型检测实现检测。该检测在定制设计的ECL检测盒中的自动化微系统中进行,该检测盒集成了流体ics、电子学和可移动光电倍增管探测器。对于大小在50至122个碱基对之间的六个靶标,获得的检测限为0.6-1.2 nmol L(-1),线性范围为1-15 nmol L(-1)。非特异性吸附和交叉反应性非常低。在单个芯片上实现了对六个靶标的检测,检测限为亚纳摩尔级。