Paper integrated aptamer-CRISPR sensing enables multiplexed small-molecule sensing in a low-cost device.
4 affiliations
- 1School of Chemical and Physical Sciences, Victoria University of Wellington Wellington 6012 New Zealand [email protected] [email protected].
- 2Independent Researcher New Zealand.
- 3Forensic Research and Development, New Zealand Institute for Public Health and Forensic Science Porirua Wellington 5022 New Zealand.
- 4Forensic Research and Development, New Zealand Institute for Public Health and Forensic Science Auckland 1025 New Zealand [email protected].
Abstract
CRISPR/Cas12a-based diagnostics achieved attomolar sensitivity for nucleic acid detection, yet their extension to small-molecule targets remains constrained by the need for solution-phase aptasensor-CRISPR coupling and laboratory infrastructure incompatible with resource-limited settings. Here we report a Paper-integrated Aptamer-Cas12a Sensing platform (PACSense) in which the complete aptasensor-CRISPR/Cas12a signal transduction cascade is air-dried within a paper substrate, enabling a one-pot, amplification-free small-molecule detection workflow that is initiated by sample addition alone. Two orthogonal sensing modalities are demonstrated: first, a signal-OFF bivalent aptamer channel for adenosine 5'-triphosphate (ATP), achieving solution- and paper-phase limits of detection of 336 nM and 446 nM respectively, and second, a signal-ON complementary strand-displacement channel for cocaine, which to our knowledge represents the first coupling of the NC195 aptamer to CRISPR/Cas12a sensing, with solution- and paper-phase LODs of 0.605 µM and 16.15 µM respectively. Both channels are operated simultaneously on a single multiplexed paper chip with a cross-channel geometry that routes sample passively to four spatially distinct reaction zones, enabling independent and simultaneous detection confirmed by fluorescence imaging under a 495 nm blue LED. Air-dried chips retain 76% of initial fluorescence signal after 30 days of ambient storage without cold chain, with a 93% signal retention measured from the functional equilibration baseline. The complete assay workflow is completed within 20 minutes using a reusable sodium acetate hand warmer for incubation and a 3D-printed smartphone-compatible reader, designed to align with ASSURED criteria for point-of-care diagnostics. PACSense establishes paper-integrated aptasensor-CRISPR sensing as a viable route to multiplexed, laboratory-instrument-free small-molecule diagnostics at the point of need.
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BibTeX
@article{sen2026paper,
title = {{Paper integrated aptamer-CRISPR sensing enables multiplexed small-molecule sensing in a low-cost device.}},
author = {Anindita Sen and Douglas James Bock and Tegan Foster and Rachel Fleming and Justin M Hodgkiss},
journal = {RSC advances},
year = {2026},
month = oct,
doi = {10.1039/d6ra08018a},
url = {https://europepmc.org/article/MED/42899138},
note = {CC BY-NC},
}
RIS
TY - JOUR
TI - Paper integrated aptamer-CRISPR sensing enables multiplexed small-molecule sensing in a low-cost device.
AU - Anindita Sen
AU - Douglas James Bock
AU - Tegan Foster
AU - Rachel Fleming
AU - Justin M Hodgkiss
PY - 2026
DA - 2026/10/19
JO - RSC advances
DO - 10.1039/d6ra08018a
UR - https://europepmc.org/article/MED/42899138
AB - CRISPR/Cas12a-based diagnostics achieved attomolar sensitivity for nucleic acid detection, yet their extension to small-molecule targets remains constrained by the need for solution-phase aptasensor-CRISPR coupling and laboratory infrastructure incompatible with resource-limited settings. Here we report a Paper-integrated Aptamer-Cas12a Sensing platform (PACSense) in which the complete aptasensor-CRISPR/Cas12a signal transduction cascade is air-dried within a paper substrate, enabling a one-pot, amplification-free small-molecule detection workflow that is initiated by sample addition alone. Two orthogonal sensing modalities are demonstrated: first, a signal-OFF bivalent aptamer channel for adenosine 5'-triphosphate (ATP), achieving solution- and paper-phase limits of detection of 336 nM and 446 nM respectively, and second, a signal-ON complementary strand-displacement channel for cocaine, which to our knowledge represents the first coupling of the NC195 aptamer to CRISPR/Cas12a sensing, with solution- and paper-phase LODs of 0.605 µM and 16.15 µM respectively. Both channels are operated simultaneously on a single multiplexed paper chip with a cross-channel geometry that routes sample passively to four spatially distinct reaction zones, enabling independent and simultaneous detection confirmed by fluorescence imaging under a 495 nm blue LED. Air-dried chips retain 76% of initial fluorescence signal after 30 days of ambient storage without cold chain, with a 93% signal retention measured from the functional equilibration baseline. The complete assay workflow is completed within 20 minutes using a reusable sodium acetate hand warmer for incubation and a 3D-printed smartphone-compatible reader, designed to align with ASSURED criteria for point-of-care diagnostics. PACSense establishes paper-integrated aptasensor-CRISPR sensing as a viable route to multiplexed, laboratory-instrument-free small-molecule diagnostics at the point of need.
SN - 2046-2069
ER -