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Microchip RT-PCR Detection of Nasopharyngeal SARS-CoV-2 Samples.
Cojocaru, Razvan; Yaseen, Iqra; Unrau, Peter J; Lowe, Christopher F; Ritchie, Gordon; Romney, Marc G; Sin, Don D; Gill, Sikander; Slyadnev, Maxim.
  • Cojocaru R; Department of Molecular Biology and Biochemistry, Simon Fraser University, Burnaby, British Columbia.
  • Yaseen I; Department of Molecular Biology and Biochemistry, Simon Fraser University, Burnaby, British Columbia.
  • Unrau PJ; Department of Molecular Biology and Biochemistry, Simon Fraser University, Burnaby, British Columbia. Electronic address: punrau@sfu.ca.
  • Lowe CF; Division of Medical Microbiology and Virology, St. Paul's Hospital, Vancouver, British Columbia; Department of Pathology and Laboratory Medicine, University of British Columbia, Vancouver, British Columbia.
  • Ritchie G; Division of Medical Microbiology and Virology, St. Paul's Hospital, Vancouver, British Columbia.
  • Romney MG; Division of Medical Microbiology and Virology, St. Paul's Hospital, Vancouver, British Columbia.
  • Sin DD; Centre for Heart Lung Innovation, St. Paul's Hospital, Vancouver, British Columbia; Department of Medicine (Respirology), University of British Columbia, Vancouver, British Columbia.
  • Gill S; Lumex Instruments Canada, Mission, British Columbia, Canada.
  • Slyadnev M; Lumex Instruments Canada, Mission, British Columbia, Canada.
J Mol Diagn ; 23(6): 683-690, 2021 06.
Article in English | MEDLINE | ID: covidwho-1121530
ABSTRACT
Fast, accurate, and reliable diagnostic tests are critical for controlling the spread of the coronavirus disease 2019 (COVID-19) associated with severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection. The current gold standard for testing is real-time PCR; however, during the current pandemic, supplies of testing kits and reagents have been limited. We report the validation of a rapid (30 minutes), user-friendly, and accurate microchip real-time PCR assay for detection of SARS-CoV-2 from nasopharyngeal swab RNA extracts. Microchips preloaded with COVID-19 primers and probes for the N gene accommodate 1.2-µL reaction volumes, lowering the required reagents by 10-fold compared with tube-based real-time PCR. We validated our assay using contrived reference samples and 21 clinical samples from patients in Canada, determining a limit of detection of 1 copy per reaction. The microchip real-time PCR provides a significantly lower resource alternative to the Centers for Disease Control and Prevention-approved real-time RT-PCR assays with comparable sensitivity, showing 100% positive and negative predictive agreement of clinical samples.
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Full text: Available Collection: International databases Database: MEDLINE Main subject: RNA, Viral / Lab-On-A-Chip Devices / Real-Time Polymerase Chain Reaction / COVID-19 Nucleic Acid Testing / SARS-CoV-2 / COVID-19 Type of study: Diagnostic study / Observational study / Prognostic study Limits: Humans Country/Region as subject: North America Language: English Journal: J Mol Diagn Journal subject: Molecular Biology Year: 2021 Document Type: Article

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Full text: Available Collection: International databases Database: MEDLINE Main subject: RNA, Viral / Lab-On-A-Chip Devices / Real-Time Polymerase Chain Reaction / COVID-19 Nucleic Acid Testing / SARS-CoV-2 / COVID-19 Type of study: Diagnostic study / Observational study / Prognostic study Limits: Humans Country/Region as subject: North America Language: English Journal: J Mol Diagn Journal subject: Molecular Biology Year: 2021 Document Type: Article