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Exp Neurobiol. 2020 Mar 11. doi: 10.5607/en20009. [Epub ahead of print]

Development of a Laboratory-safe and Low-cost Detection Protocol for SARS-CoV-2 of the Coronavirus Disease 2019 (COVID-19).

Author information

1
Center for Cognition and Sociality, Cognitive Glioscience Group, Institute for Basic Science, Daejeon 34126, Korea.
2
Department of Biological Sciences, Korea Advanced Institute of Science and Technology (KAIST), Daejeon 34141, Korea.
3
KU-KIST Graduate School of Converging Science and Technology, Korea University, Seoul 02841, Korea.
4
Department of Otorhinolaryngology, Seoul National University Bundang Hospital, Seongnam 13620, Korea.
5
Center for RNA Research, Institute for Basic Science, Seoul 08826, Korea.
6
School of Biological Sciences, Seoul National University, Seoul 08826, Korea.

Abstract

The severe acute respiratory coronavirus 2 (SARS-CoV-2), which emerged in December 2019 in Wuhan, China, has spread rapidly to over a dozen countries. Especially, the spike of case numbers in South Korea sparks pandemic worries. This virus is reported to spread mainly through personto- person contact via respiratory droplets generated by coughing and sneezing, or possibly through surface contaminated by people coughing or sneezing on them. More critically, there have been reports about the possibility of this virus to transmit even before a virus-carrying person to show symptoms. Therefore, a low-cost, easy-access protocol for early detection of this virus is desperately needed. Here, we have established a real-time reverse-transcription PCR (rtPCR)-based assay protocol composed of easy specimen self-collection from a subject via pharyngeal swab, Trizolbased RNA purification, and SYBR Green-based rtPCR. This protocol shows an accuracy and sensitivity limit of 1-10 virus particles as we tested with a known lentivirus. The cost for each sample is estimated to be less than 15 US dollars. Overall time it takes for an entire protocol is estimated to be less than 4 hours. We propose a cost-effective, quick-and-easy method for early detection of SARS-CoV-2 at any conventional Biosafety Level II laboratories that are equipped with a rtPCR machine. Our newly developed protocol should be helpful for a first-hand screening of the asymptomatic virus-carriers for further prevention of transmission and early intervention and treatment for the rapidly propagating virus.

KEYWORDS:

COVID-19; Communicable diseases; Coronavirus; Diagnostic techniques and procedures; Emerging; Infectious disease; SARS virus

PMID:
32156101
DOI:
10.5607/en20009
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