Electrochemiluminescence DNA sensor array for multiplex detection of biowarfare agents

Abstract

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.

Electrochemiluminescence Biosensors Using Screen-Printed Electrodes

“…For this reason, the development of multiplex ECL assays for biowarfare agent is of great interest. O´Sullivan et al [ 173 ] reported the develop of a multiplex detection ECL DNA sensor for the simultaneous detection of six pathogens: Brucella, melitensis, Bacillus anthracis, Burkholderia mallei, Francisella tularensis, Bacillus thuringiensis var. kurstaki and Coxiella burnetiid .…”

Advance detection technologies for select biothreat agents

“…DNA array-based sensors: The DNA array class of detectors relies on comparing the DNA taken from microorganisms in a sample with the known DNA of known biowarfare agents. Researchers have in recent years been sequencing the DNA of an array of potential agents, using common gene sequencing methods (ESpehar-Délèze et al, 2015).…”

Recent advances in electrogenerated chemiluminescence biosensing methods for pharmaceuticals

“…Aptamers as molecular recognition substances appear to be excellent alternatives to antibodies due to their ease of production in vitro, wide target range, modification ease, reversible thermal denaturation, and unlimited shelf life. Recently, a number of ECL aptamer-based biosensors for the determination of a small molecule drug have been designed employing aptamers as molecular recognition elements and Ru(II) complex served as an ECL label [68] , [69] , [70] , [71] , [72] , [73] , as listed in Table 3 .…”

“… Drugs/analytes Systems Linear ranges LOD (S/N = 3) Samples Ref. Cocaine Signal-on, Ru1-tagged aptamer/4-ABSA/ PIGE in 0.10 M PBS-0.10 M TPA (pH 7.40) 50 pM to 5 nM 10 pM – [68] Cocaine Capture apt-probe assembled on Au electrode, signal apt-probe labeled with RuSiNPs in PBS (pH7.4)-1.0 M dibutylamino-ethanol 0.01–1.0 nM 3.7 pM Banknotes [69] Adenosine Sandwich type, Au electrode, Ru(phen) 3 2+ as interactor 0.5–7 nM 0.15 nM – [70] Adenosine Signal off, Ru1-tagged aptamers/SWCNTs/GCE 0.1–500 nM 0.05 nM – [71] Malachite green (MG) Chloramphenicol (CAP) Aptasensor array using CdS QDs and luminol-AuNPs as labels, 0.1 M PBS (pH 8.0)-12 mM H 2 O 2 0.1–100 nM MG 0.03 nM MG Fish samples [72] 0.2–150 nM CAP 0.07 nM CAP Biowarfare agents SPC electrode array (ss-DNA), Ru(bpy) 3 2+ -labeled DNA reporter, sandwich-type hybridisation assay in 300 mM PB-100 mM TPA-0.02% Triton X-100 (pH 7.6) 0–16 nM 0.6–1.2 nM for six targets – [73] …”