High Sensitivity, Rapid Detection of Virus in High Traffic Environments

Lauren Waller; Zhilin Guo; Rui Tang; Zunming Zhang; Edward Wang; Jarred Yasuhara-Bell; Louise Laurent; Yu-Hwa Lo

doi:10.3389/fbioe.2022.877603

High Sensitivity, Rapid Detection of Virus in High Traffic Environments

Front Bioeng Biotechnol. 2022 Mar 24:10:877603. doi: 10.3389/fbioe.2022.877603. eCollection 2022.

Authors

Lauren Waller¹, Zhilin Guo², Rui Tang³, Zunming Zhang³, Edward Wang², Jarred Yasuhara-Bell⁴, Louise Laurent⁵, Yu-Hwa Lo³

Affiliations

¹ Department of Bioengineering, University of California, San Diego, San Diego, CA, United States.
² Department of Mechanical and Aerospace Engineering, University of California, San Diego, San Diego, CA, United States.
³ Department of Electrical and Computer Engineering, University of California, San Diego, San Diego, CA, United States.
⁴ Independent Consultant, San Diego, CA, United States.
⁵ Department of Obstetrics and Gynecology and Reproductive Sciences, University of California, San Diego, San Diego, CA, United States.

Abstract

The global pandemic caused by the SARS-CoV-2 virus has underscored the need for rapid, simple, scalable, and high-throughput multiplex diagnostics in non-laboratory settings. Here we demonstrate a multiplex reverse-transcription loop-mediated isothermal amplification (RT-LAMP) coupled with a gold nanoparticle-based lateral flow immunoassay (LFIA) capable of detecting up to three unique viral gene targets in 15 min. RT-LAMP primers associated with three separate gene targets from the SARS-CoV-2 virus (Orf1ab, Envelope, and Nucleocapsid) were added to a one-pot mix. A colorimetric change from red to yellow occurs in the presence of a positive sample. Positive samples are run through a LFIA to achieve specificity on a multiplex three-test line paper assay. Positive results are indicated by a characteristic crimson line. The device is almost fully automated and is deployable in any community setting with a power source.

Keywords: COVID19; RT-LAMP; SARS-cov-2; lateral flow assay; multiplexed loop mediated isothermal amplification; point of care diagnostics; rapid diagnostics.