US CDC Real-Time Reverse Transcription PCR Panel for Detection of Severe Acute Respiratory Syndrome Coronavirus 2

• Published in: Emerging infectious diseases Vol. 26; no. 8; pp. 1654 - 1665
• Main Authors: Lu, Xiaoyan, Wang, Lijuan, Sakthivel, Senthilkumar K., Whitaker, Brett, Murray, Janna, Kamili, Shifaq, Lynch, Brian, Malapati, Lakshmi, Burke, Stephen A., Harcourt, Jennifer, Tamin, Azaibi, Thornburg, Natalie J., Villanueva, Julie M., Lindstrom, Stephen
• Format: Journal Article
• Language: English
• Published: United States U.S. National Center for Infectious Diseases 01.08.2020; Centers for Disease Control and Prevention
• Subjects: 2019 novel coronavirus disease; Betacoronavirus - genetics; Biomarkers - analysis; Centers for Disease Control and Prevention, U.S; coronavirus disease; Coronavirus Infections - diagnosis; Coronavirus Infections - virology; Coronavirus Nucleocapsid Proteins; Coronaviruses; COVID-19; Deoxyribonucleic acid; Disease control; DNA; DNA Primers - chemical synthesis; DNA Primers - genetics; Feces - virology; Fluoresceins - chemistry; Fluorescent Dyes - chemistry; Genetic aspects; Genomes; Health aspects; Humans; Limit of Detection; Medical laboratories; Nasopharynx - virology; Nucleocapsid Proteins - genetics; Pandemics; Phosphoproteins; Pneumonia, Viral - diagnosis; Pneumonia, Viral - virology; Polymerase chain reaction; Real-Time Polymerase Chain Reaction - methods; Real-Time Polymerase Chain Reaction - standards; Reproducibility of Results; Respiratory diseases; RNA; RNA polymerase; RNA, Viral - genetics; SARS-CoV-2; Severe acute respiratory syndrome coronavirus 2; Sputum - virology; Transcription (Genetics); United States; US CDC Real-Time Reverse Transcription PCR Panel for Detection of Severe Acute Respiratory Syndrome Coronavirus 2; Viruses; United States; United States > US; China; COVID-19; SARS-CoV-2; viruses; severe acute respiratory syndrome coronavirus 2; zoonoses; 2019 novel coronavirus disease; coronavirus disease; real-time RT-PCR; real-time reverse transcription PCR; respiratory infections
• ISSN: 1080-6040; 1080-6059; 1080-6059
• DOI: 10.3201/eid2608.201246

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) was identified as the etiologic agent associated with coronavirus disease, which emerged in late 2019. In response, we developed a diagnostic panel consisting of 3 real-time reverse transcription PCR assays targeting the nucleocapsid gene and evaluated use of these assays for detecting SARS-CoV-2 infection. All assays demonstrated a linear dynamic range of 8 orders of magnitude and an analytical limit of detection of 5 copies/reaction of quantified RNA transcripts and 1 x 10 50% tissue culture infectious dose/mL of cell-cultured SARS-CoV-2. All assays performed comparably with nasopharyngeal and oropharyngeal secretions, serum, and fecal specimens spiked with cultured virus. We obtained no false-positive amplifications with other human coronaviruses or common respiratory pathogens. Results from all 3 assays were highly correlated during clinical specimen testing. On February 4, 2020, the Food and Drug Administration issued an Emergency Use Authorization to enable emergency use of this panel.