Smartphone clip-on instrument and microfluidic processor for rapid sample-to-answer detection of Zika virus in whole blood using spatial RT-LAMP

Weijing Wang; Hankeun Lee; Aaron M. Jankelow; Trung-Hieu Hoang; Amanda Bacon; Fu Sun; Seol Chae; Victoria Kindratenko; Katherine Koprowski; Robert A. Stavins; Dylann Ceriani; Zachary Engelder; William P. King; Minh N. Do; Rashid Bashir; Enrique Valera; Brian T. Cunningham

doi:10.1117/12.2649267

16 March 2023 Smartphone clip-on instrument and microfluidic processor for rapid sample-to-answer detection of Zika virus in whole blood using spatial RT-LAMP

Weijing Wang, Hankeun Lee, Aaron M. Jankelow, Trung-Hieu Hoang, Amanda Bacon, Fu Sun, Seol Chae, Victoria Kindratenko, Katherine Koprowski, Robert A. Stavins, Dylann Ceriani, Zachary Engelder, William P. King, Minh N. Do, Rashid Bashir, Enrique Valera, Brian T. Cunningham

Author Affiliations +

Proceedings Volume 12387, Optical Diagnostics and Sensing XXIII: Toward Point-of-Care Diagnostics; 123870D (2023) https://doi.org/10.1117/12.2649267
Event: SPIE BiOS, 2023, San Francisco, California, United States

Abstract

We report an integrated system for rapid sample-to-answer detection of a viral pathogen in a droplet of whole blood comprised of a two-stage microfluidic cartridge for sample processing and nucleic acid amplification, and a clip-on detection instrument that interfaces with the image sensor of a smartphone. The cartridge is designed to release RNA from the Zika virus in whole blood using chemical lysis, followed by mixing with the assay buffer for performing reverse-transcriptase loop-mediated isothermal amplification (RT-LAMP) reactions in six parallel microfluidic compartments. The battery-powered instrument heats the compartments from below, while LEDs illuminate from above. Fluorescence generation in the compartments is dynamically monitored by a smartphone camera. We characterize the assay time and detection limits for detecting Zika RNA and gamma-irradiated Zika virus spiked into buffer and whole blood and compare the performance of the same assay when conducted in conventional PCR tubes. Our approach for kinetic monitoring of the fluorescence-generating process in the microfluidic compartments enables spatial analysis of early fluorescent “bloom” events for positive samples. We show that dynamic image analysis reduces the time required to designate an assay as a positive test to 22 minutes, compared to ~30-45 minutes for conventional analysis of the average fluorescent intensity of the entire compartment. We achieve a total sample-to-answer time in the range of 17-32 minutes, while demonstrating a viral RNA detection as low as 2.70×10² copies/ul, and a gamma-irradiated virus of 103 virus particles in a single 12.5 microliter droplet blood sample.

Conference Presentation

Citation Download Citation

Weijing Wang, Hankeun Lee, Aaron M. Jankelow, Trung-Hieu Hoang, Amanda Bacon, Fu Sun, Seol Chae, Victoria Kindratenko, Katherine Koprowski, Robert A. Stavins, Dylann Ceriani, Zachary Engelder, William P. King, Minh N. Do, Rashid Bashir, Enrique Valera, and Brian T. Cunningham "Smartphone clip-on instrument and microfluidic processor for rapid sample-to-answer detection of Zika virus in whole blood using spatial RT-LAMP", Proc. SPIE 12387, Optical Diagnostics and Sensing XXIII: Toward Point-of-Care Diagnostics, 123870D (16 March 2023); https://doi.org/10.1117/12.2649267

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