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Sputum Processing Method for Lateral Flow Immunochromatographic Assays to Detect Coronaviruses

  • Aram Kang (College of Pharmacy, Korea University) ;
  • Minjoo Yeom (College of Pharmacy, Korea University) ;
  • Hyekwon Kim (Department of Microbiology, Chungbuk National University) ;
  • Sun-Woo Yoon (Korea Research Institute of Bioscience and Biotechnology) ;
  • Dae-Gwin Jeong (Korea Research Institute of Bioscience and Biotechnology) ;
  • Hyong-Joon Moon (College of Healthcare & Biotechnology, Semyung University) ;
  • Kwang-Soo Lyoo (Korea Zoonosis Research Institute, Chonbuk National University) ;
  • Woonsung Na (College of Veterinary Medicine, Chonnam National University) ;
  • Daesub Song (College of Pharmacy, Korea University)
  • Received : 2020.10.04
  • Accepted : 2021.01.30
  • Published : 2021.02.28
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Abstract

Coronavirus causes an infectious disease in various species and crosses the species barriers leading to the outbreak of zoonotic diseases. Due to the respiratory diseases are mainly caused in humans and viruses are replicated and excreted through the respiratory tract, the nasal fluid and sputum are mainly used for diagnosis. Early diagnosis of coronavirus plays an important role in preventing its spread and is essential for quarantine policies. For rapid decision and prompt triage of infected host, the immunochromatographic assay (ICA) has been widely used for point of care testing. However, when the ICA is applied to an expectorated sputum in which antigens are present, the viscosity of sputum interferes with the migration of the antigens on the test strip. To overcome this limitation, it is necessary to use a mucolytic agent without affecting the antigens. In this study, we combined known mucolytic agents to lower the viscosity of sputum and applied that to alpha and beta coronavirus, porcine epidemic diarrhea virus (PEDV) and Middle East respiratory syndrome coronavirus (MERS-CoV), respectively, spiked in sputum to find optimal pretreatment conditions. The pretreatment method using tris(2-carboxyethyl)phosphine (TCEP) and BSA was suitable for ICA diagnosis of sputum samples spiked with PEDV and MERS-CoV. This sensitive assay for the detection of coronavirus in sputum provides an useful information for the diagnosis of pathogen in low respiratory tract.

Keywords

Acknowledgement

This work was supported by a Korea University grant.

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