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07 20, 2021;11 (1): 14817.

Validation of real-time RT-PCR for detection of SARS-CoV-2 in the early stages of the COVID-19 outbreak in the Republic of Korea.

Yoon-Seok Chung, Nam-Joo Lee, Sang Hee Woo, Jeong-Min Kim, Heui Man Kim, Hye Jun Jo, Ye Eun Park, Myung-Guk Han
Author Information
  1. Yoon-Seok Chung: Division of Infectious Disease Diagnosis Control, Honam Regional Centers for Disease Control and Prevention, Korea Diseases Control and Prevention Agency, Gwangju-si, 61947, Republic of Korea.
  2. Nam-Joo Lee: Division of Emerging Infectious Diseases, Bureau of Infectious Disease Diagnosis Control, Korea Diseases Control and Prevention Agency, Cheongju-si, 28159, Republic of Korea.
  3. Sang Hee Woo: Division of Emerging Infectious Diseases, Bureau of Infectious Disease Diagnosis Control, Korea Diseases Control and Prevention Agency, Cheongju-si, 28159, Republic of Korea.
  4. Jeong-Min Kim: Division of Emerging Infectious Diseases, Bureau of Infectious Disease Diagnosis Control, Korea Diseases Control and Prevention Agency, Cheongju-si, 28159, Republic of Korea.
  5. Heui Man Kim: Division of Emerging Infectious Diseases, Bureau of Infectious Disease Diagnosis Control, Korea Diseases Control and Prevention Agency, Cheongju-si, 28159, Republic of Korea.
  6. Hye Jun Jo: Division of Emerging Infectious Diseases, Bureau of Infectious Disease Diagnosis Control, Korea Diseases Control and Prevention Agency, Cheongju-si, 28159, Republic of Korea.
  7. Ye Eun Park: Division of Laboratory Diagnosis Management, Bureau of Infectious Disease Diagnosis Control, Korea Diseases Control and Prevention Agency, Cheongju-si, 28159, Republic of Korea.
  8. Myung-Guk Han: Division of Viral Diseases, Bureau of Infectious Disease Diagnosis Control, Korea Diseases Control and Prevention Agency, 187 Osongsaengmyeong2-ro, Osong-eup, Heungdeok-gu, Cheongju-si, 28159, Republic of Korea. mghan@korea.kr.
PMID: 34285290 DOI: 10.1038/s41598-021-94196-3

Abstract

A real-time reverse transcription polymerase chain reaction (RT-qPCR) assay that does not require Emergency Use Authorization (EUA) reagents was tested and validated for the detection of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) during the early stages of the outbreak of coronavirus disease 2019 (COVID-19) in the Republic of Korea. Early diagnosis of COVID-19 enables timely treatment and the implementation of public health measures. We validated the sensitivity, specificity, precision, linearity, accuracy, and robustness of the RT-qPCR assay for SARS-CoV-2 detection and compared its performance with that of several EUA-approved kits. Our RT-qPCR assay was highly specific for SARS-CoV-2 as demonstrated by not amplifying 13 other viruses that cause respiratory diseases. The assay showed high linearity using a viral isolate from a patient with known COVID-19 as well as plasmids containing target SARS-CoV-2 genes as templates. The assay showed good repeatability and reproducibility with a coefficient of variation of 3%, and a SARS-CoV-2 limit of detection of 1 PFU/mL. The RT-qPCR-based assay is highly effective and can facilitate the early diagnosis of COVID-19 without the use of EUA-approved kits or reagents in the Republic of Korea.

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Grants

  1. [grant number 4800-4837-301]/This work was supported by Korea Centers for Disease Control and Prevention .
  2. [grant number 4800-4837-301]/This work was supported by Korea Centers for Disease Control and Prevention .
  3. [grant number 4800-4837-301]/This work was supported by Korea Centers for Disease Control and Prevention .
  4. [grant number 4800-4837-301]/This work was supported by Korea Centers for Disease Control and Prevention .
  5. [grant number 4800-4837-301]/This work was supported by Korea Centers for Disease Control and Prevention .
  6. [grant number 4800-4837-301]/This work was supported by Korea Centers for Disease Control and Prevention .
  7. [grant number 4800-4837-301]/This work was supported by Korea Centers for Disease Control and Prevention .
  8. [grant number 4800-4837-301]/This work was supported by Korea Centers for Disease Control and Prevention .

MeSH Term

Animals
COVID-19
COVID-19 Nucleic Acid Testing
Chlorocebus aethiops
Humans
Limit of Detection
Real-Time Polymerase Chain Reaction
Reproducibility of Results
Republic of Korea
Reverse Transcriptase Polymerase Chain Reaction
Vero Cells
Journal Article Research Support, Non-U.S. Gov't Validation Study
Article has an altmetric score of 20

Word Cloud

Created with Highcharts 10.0.0assaySARS-CoV-2COVID-19detectionRT-qPCRearlyRepublicKoreareal-timereagentsvalidatedrespiratorycoronavirusstagesoutbreakdiagnosislinearityEUA-approvedkitshighlyshowedreversetranscriptionpolymerasechainreactionrequireEmergencyUseAuthorizationEUAtestedsevereacutesyndrome2disease2019Earlyenablestimelytreatmentimplementationpublichealthmeasuressensitivityspecificityprecisionaccuracyrobustnesscomparedperformanceseveralspecificdemonstratedamplifying13virusescausediseaseshighusingviralisolatepatientknownwellplasmidscontainingtargetgenestemplatesgoodrepeatabilityreproducibilitycoefficientvariation3%limit1PFU/mLRT-qPCR-basedeffectivecanfacilitatewithoutuseValidationRT-PCR

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