期刊
ANALYTICAL CHEMISTRY
卷 93, 期 42, 页码 14187-14195出版社
AMER CHEMICAL SOC
DOI: 10.1021/acs.analchem.1c02900
关键词
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资金
- European Research Council [715268]
- Knut and Alice Wallenberg Foundation
The 3D microfluidic paper-based electrochemical NAAT presented in this study combines sensitive detection of target DNA with the integration of thread electrodes within paper devices, allowing for time-sequenced reactions. This innovative approach showed high sensitivity and specificity in detecting toxic microalgae within a short period of 95 minutes.
Nucleic acid amplification tests (NAATs) are very sensitive and specific methods, but they mainly rely on centralized laboratories and therefore are not suitable for point-of-care testing. Here, we present a 3D microfluidic paper-based electrochemical NAAT. These devices use off-the-shelf gold plasma-coated threads to integrate electroanalytical readouts using ex situ self-assembled monolayer formation on the threads prior to assembling into the paper device. They further include a sandwich hybridization assay with sample incubation, rinsing, and detection steps all integrated using movable stacks of filter papers to allow time-sequenced reactions. The devices use glass fiber substrates for storing recombinase polymerase amplification reagents and conducting the isothermal amplification. We used the paper-based device for the detection of the toxic microalgae Ostreopsis cf. ovata. The NAAT, completed in 95 min, attained a limit of detection of 0.06 pM target synthetic DNA and was able to detect 1 ng/mu L O. cf. ovata genomic DNA with negligible cross-reactivity from a closely related microalgae species. We think that the integration of thread electrodes within paper-based devices paves the way for digital one-time use NAATs and numerous other advanced electroanalytical paper- or textile-based devices.
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