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Updated: Oct 13, 2025
Multiplexed Isothermal Amplification Based Diagnostic Platform to Detect Zika, Chikungunya, and Dengue 1
Published on: March 13, 2018
A Valve-Enabled Sample Preparation Device with Isothermal Amplification for Multiplexed Virus Detection at the
Carlos Manzanas1, Md Mahbubul Alam2,3, Julia C Loeb2,3
1Interdisciplinary Microsystems Group, Department of Mechanical and Aerospace Engineering, University of Florida, P. O. Box 116250, Gainesville, Florida 32611, United States.
Abstract:
Early and accurate detection of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) and influenza viruses at the point-of-care is crucial for reducing disease transmission during the current pandemic and future flu seasons. To prepare for potential cocirculation of these two viruses, we report a valve-enabled, paper-based sample preparation device integrated with isothermal amplification for their simultaneous detection. The device incorporates (1) virus lysis and RNA enrichment, enabled by ball-based valves for sequential delivery of reagents with no pipet requirement, (2) reverse transcription loop-mediated isothermal amplification, carried out in a coffee mug, and (3) colorimetric detection. We have used the device for simultaneously detecting inactivated SARS-CoV-2 and influenza A H1N1 viruses in 50 min, with limits of detection at 2 and 6 genome equivalents, respectively. The device was further demonstrated to detect both viruses in environmental samples.
