Electrochemiluminescence DNA sensor array for multiplex detection of biowarfare agents.

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.

Electrochemiluminescent DNA sensor based on controlled Zn-mediated grafting of diazonium precursors.

Controlled Zn-mediated grafting of a thin layer of a diazonium salt was used to functionalise a carbon electrode with ruthenium(II)-tris-bipyridine (Ru)-labelled DNA for use as a capture probe in an electrochemiluminescent genosensor. A secondary reporter probe was labelled with a ferrocene (Fc) molecule, and in the presence of the single-stranded DNA target a genocomplex formed, where the Fc-label effectively quenched the electrochemiluminescence of the signal emitted from the Ru-label. The spacing of the labels for maximum sensitivity and minimum detection limit was optimised, and the signal reproducibility and stability of the method was established.

Potentiometric strip cell based on carbon nanotubes as transducer layer: toward low-cost decentralized measurements.

In this study, we developed a potentiometric planar strip cell based on single-walled carbon nanotubes that aims to exploit the attributes of solid-contact ion-selective electrodes for decentralized measurements. That is, the ion-selective and reference electrodes have been simultaneously miniaturized onto a plastic planar substrate by screen-printing and drop-casting techniques, obtaining disposable strip cells with satisfactory performance characteristics (i.e., the sensitivity is 57.4 ± 1.3 mV/dec, the response time is ≤30 s within the linear range from log a(K+) = -5 to -2, and the limit of detection is -6.5), no need of maintenance during long dry storage, quick signal stabilization, and light insensitivity in short-term measurements. We also show how the new potentiometric strip cell makes it possible to perform decentralized and rapid determinations of ions in real samples, such as saliva or beverages.

Disposable planar reference electrode based on carbon nanotubes and polyacrylate membrane.

In this technical note, we report a new all-solid-state planar reference electrode based on single-walled carbon nanotubes and photocured poly(n-butylacrylate) (poly(nBA)) membrane containing the Ag/AgCl/Cl(-) ion system. Single-walled carbon nanotubes functionalized with octadecylamide (SWCNT-ODA) and deposited by drop-casting onto a disposable screen-printed electrode are an excellent all-solid-state transducer. The novel potentiometric planar reference electrode shows low potential variability (calibration slopes inferior to 2 mV/dec) for a wide range of chemical species (i.e., ions, small molecules, proteins) in a wide calibration range, redox pairs, changes in pH, and changes in ambient light. Potentiometric medium-term signal stability (-0.9 ± 0.2 mV/h) and electrochemical impedance characterization confirm the correct solid contact between the SWCNT-ODA layer and photocured poly(nBA) membrane. Overall, the materials used and the simple fabrication by screen-printing and drop-casting enable a high throughput and highly parallel and cost-effective mass manufacture of the new disposable reference electrode. Moreover, the reference electrode has a long shelf life, a characteristic that can be of special interest in decentralized and multiplexing potentiometric analysis.

Screen-printed microsystems for the ultrasensitive electrochemical detection of alkaline phosphatase.

Screen printing technique has been used to manufacture a microsystem where the graphite-based electrodes hold both a functional and an architectural task. The thick film manufacturing technique has proved valid to develop a very low volume (ca. 20 microL) device where different electrochemical operations can be very efficiently performed. Biomolecule immobilisation within the microsystem for biosensors applications has been explored by inducing and optimizing the in situ generation of a potential pulse polypyrrole electropolymerised film entrapping either glucose oxidase or glucose dehydrogenase. This biomodified microsystem was applied to the ultrasensitive electrochemical detection of alkaline phosphatase yielding limits of detection below 10(-12) M for glucose oxidase and of 10(-15) M for glucose dehydrogenase modified systems, within 15 min of incubation time. The results obtained showed the advantages of using low volume microsystems in combination with an optimised polypyrrole-enzyme film, which displayed a good immobilisation efficiency in conjunction with a good diffusion of species through. Ultrasensitive detection of AP in combination with a stable and reproducible surface modification for entrapping of biomolecules opens the window for new electrochemical detection platform with great potential for integrated biosensor applications.