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

Research output: Chapter in Book/Report/Conference proceedingConference contribution

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 2-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 2.70x102 copies/ul, and a gamma-irradiated virus of 103 virus particles in a single 12.5 microliter droplet blood sample.

Original languageEnglish (US)
Title of host publicationOptical Diagnostics and Sensing XXIII
Subtitle of host publicationToward Point-of-Care Diagnostics
EditorsGerard L. Cote
PublisherSPIE
ISBN (Electronic)9781510658790
DOIs
StatePublished - 2023
EventOptical Diagnostics and Sensing XXIII: Toward Point-of-Care Diagnostics 2023 - San Francisco, United States
Duration: Jan 30 2023Jan 31 2023

Publication series

NameProgress in Biomedical Optics and Imaging - Proceedings of SPIE
Volume12387
ISSN (Print)1605-7422

Conference

ConferenceOptical Diagnostics and Sensing XXIII: Toward Point-of-Care Diagnostics 2023
Country/TerritoryUnited States
CitySan Francisco
Period1/30/231/31/23

Keywords

  • Fluorescence
  • Fluorescence
  • Microfluidic chip
  • Nucleic acid detection
  • Point of care
  • RT-LAMP
  • Smartphone-based device
  • Zika virus

ASJC Scopus subject areas

  • Electronic, Optical and Magnetic Materials
  • Atomic and Molecular Physics, and Optics
  • Biomaterials
  • Radiology Nuclear Medicine and imaging

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