A SARS-CoV-2 coronavirus nucleocapsid protein antigen-detecting lateral flow assay

PLoS One. 2021 Nov 10;16(11):e0258819. doi: 10.1371/journal.pone.0258819. eCollection 2021.

Abstract

Inexpensive, simple, rapid diagnostics are necessary for efficient detection, treatment, and mitigation of COVID-19. Assays for SARS-CoV2 using reverse transcription polymerase chain reaction (RT-PCR) offer good sensitivity and excellent specificity, but are expensive, slowed by transport to centralized testing laboratories, and often unavailable. Antigen-based assays are inexpensive and can be rapidly mass-produced and deployed at point-of-care, with lateral flow assays (LFAs) being the most common format. While various manufacturers have produced commercially available SARS-Cov2 antigen LFAs, access to validated tests remains difficult or cost prohibitive in low-and middle-income countries. Herein, we present a visually read open-access LFA (OA-LFA) using commercially-available antibodies and materials for the detection of SARS-CoV-2. The LFA yielded a Limit of Detection (LOD) of 4 TCID50/swab of gamma irradiated SARS-CoV-2 virus, meeting the acceptable analytical sensitivity outlined by in World Health Organization target product profile. The open-source architecture presented in this manuscript provides a template for manufacturers around the globe to rapidly design a SARS-CoV2 antigen test.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Antigens, Viral / immunology*
  • COVID-19 / diagnosis*
  • COVID-19 / immunology*
  • COVID-19 / virology
  • COVID-19 Testing / methods*
  • Coronavirus Nucleocapsid Proteins / immunology*
  • Humans
  • Limit of Detection
  • Point-of-Care Systems
  • RNA, Viral / immunology
  • SARS-CoV-2 / immunology*
  • Sensitivity and Specificity

Substances

  • Antigens, Viral
  • Coronavirus Nucleocapsid Proteins
  • RNA, Viral

Grants and funding

Funding provided by The Global Good Fund and Global Health Labs, a nonprofit organization created by Gates Ventures and the Gates Foundation to develop innovative solutions to address unmet needs in primary health care centers and the last mile.