Development of a Laboratory-safe and Low-cost Detection Protocol for SARS-CoV-2 of the Coronavirus Disease 2019 (COVID-19)
Authors: Dongwan Kim
Hyeshik Chang
Solji Lee
C Justin Lee
Joungha Won
Tai Young Kim
Myungsun Park
Byung Yoon Choi
V Narry Kim
Mingu Gordon Park
Journal: Experimental neurobiology (2020)
Discipline: NEUROSCIENCE & BEHAVIOR
Keywords: COVID-19; Communicable diseases; Coronavirus; Diagnostic techniques and procedures; Emerging
Affiliations: Center for Cognition and Sociality, Cognitive Glioscience Group, Institute for Basic Science, Daejeon 34126, Korea., Department of Biological Sciences, Korea Advanced Institute of Science and Technology (KAIST), Daejeon 34141, Korea., KU-KIST Graduate School of Converging Science and Technology, Korea University, Seoul 02841, Korea., Department of Otorhinolaryngology, Seoul National University Bundang Hospital, Seongnam 13620, Korea., Center for RNA Research, Institute for Basic Science, Seoul 08826, Korea., 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 person-to-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, Trizol-based 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. 

Retracted: No

PubPeer comments:
 **The second paragraph of this article says:**  Of the 126 Germans sent to Germany from Wuhan, China, on February 1, 2020, 114 patients were asymptomatic but found to be positive for SARS-CoV-2, suggesting that a symptom-based screening process may not be sufficiently effective [4] **However, [the cited letter] says:**  In addition to the preplanned multistep process of screening for signs and symptoms of infection and observing the asymptomatic cohort in quarantine, we decided to offer a throat swab to test for SARS-CoV-2 in each of the 115 travelers who had passed triage. A total of 114 passengers consented to the test. Two of the 114 persons (1.8%) in this cohort of travelers who had passed the symptoms-based screening tested positive for SARS-CoV-2 by RT-PCR (cycle threshold value in the two samples, 24.39 and 30.25, respectively). Testing with a second protocol consisting of two commercial sets (LightMix Modular SARS and Wuhan CoV E-gene, and LightMix Modular Wuhan CoV RdRP-gene, both produced by TIB MOLBIOL) and retesting of the positive samples at the Institute of Virology, Philipps University Marburg, in Marburg, Germany, confirmed the results. In addition, the isolation of SARS-CoV-2 from both samples in cell culture of Caco-2 cells indicated potential infectivity (see the Supplementary Appendix, available with the full text of this letter at NEJM.org). **Or more briefly:** 114 passengers were tested, but only two tested positive for SARS-CoV-2. The authors appear to have misread the letter they re citing. ,