Development of a qualitative real-time RT-PCR assay for the detection of SARS-CoV-2: a guide and case study in setting up an emergency-use, laboratory-developed molecular microbiological assay.

Autor:	Anahtar MN; Department of Pathology and Harvard Medical School, Massachusetts General Hospital, Boston, Massachusetts, USA., Shaw BM; Infectious Disease and Microbiome Program, Eli and Edythe L. Broad Institute of Harvard and MIT, Cambridge, Massachusetts, USA.; Department of Medicine, Division of Infectious Diseases, Massachusetts General Hospital, Boston, Massachusetts, USA., Slater D; Department of Medicine, Division of Infectious Diseases, Massachusetts General Hospital, Boston, Massachusetts, USA., Byrne EH; Infectious Disease and Microbiome Program, Eli and Edythe L. Broad Institute of Harvard and MIT, Cambridge, Massachusetts, USA., Botti-Lodovico Y; Infectious Disease and Microbiome Program, Eli and Edythe L. Broad Institute of Harvard and MIT, Cambridge, Massachusetts, USA., Adams G; Infectious Disease and Microbiome Program, Eli and Edythe L. Broad Institute of Harvard and MIT, Cambridge, Massachusetts, USA.; Department of Medicine, Division of Infectious Diseases, Massachusetts General Hospital, Boston, Massachusetts, USA., Schaffner SF; Infectious Disease and Microbiome Program, Eli and Edythe L. Broad Institute of Harvard and MIT, Cambridge, Massachusetts, USA.; Center for Systems Biology, Department of Organismic and Evolutionary Biology, Harvard University, Cambridge, Massachusetts, USA., Eversley J; Department of Pathology and Harvard Medical School, Massachusetts General Hospital, Boston, Massachusetts, USA., McGrath GEG; Department of Medicine, Division of Infectious Diseases, Massachusetts General Hospital, Boston, Massachusetts, USA., Gogakos T; Department of Pathology and Harvard Medical School, Massachusetts General Hospital, Boston, Massachusetts, USA., Lennerz J; Department of Pathology and Harvard Medical School, Massachusetts General Hospital, Boston, Massachusetts, USA., Marble HD; Department of Pathology and Harvard Medical School, Massachusetts General Hospital, Boston, Massachusetts, USA., Ritterhouse LL; Department of Pathology and Harvard Medical School, Massachusetts General Hospital, Boston, Massachusetts, USA., Batten JM; Department of Pathology and Harvard Medical School, Massachusetts General Hospital, Boston, Massachusetts, USA., Georgantas NZ; Department of Pathology and Harvard Medical School, Massachusetts General Hospital, Boston, Massachusetts, USA., Pellerin R; Department of Pathology and Harvard Medical School, Massachusetts General Hospital, Boston, Massachusetts, USA., Signorelli S; Department of Pathology and Harvard Medical School, Massachusetts General Hospital, Boston, Massachusetts, USA., Thierauf J; Department of Pathology and Harvard Medical School, Massachusetts General Hospital, Boston, Massachusetts, USA.; Department of Otorhinolaryngology, University Hospital Heidelberg, Heidelberg, Baden-Württemberg, Germany., Kemball M; Infectious Disease and Microbiome Program, Eli and Edythe L. Broad Institute of Harvard and MIT, Cambridge, Massachusetts, USA.; Center for Systems Biology, Department of Organismic and Evolutionary Biology, Harvard University, Cambridge, Massachusetts, USA., Happi C; Department of Biological Sciences, Redeemer's University, Ede, Osun, Nigeria.; African Center of Excellence for Genomics of Infectious Diseases, Redeemer's University, Ede, Osun, Nigeria., Grant DS; Viral Hemorrhagic Fever Program, Kenema Government Hospital, Kenema, Sierra Leone.; College of Medicine and Allied Health Sciences, University of Sierra Leone, Freetown, Sierra Leone., Ndiaye D; African Center of Excellence for Genomics of Infectious Diseases, Redeemer's University, Ede, Osun, Nigeria.; Department of Mycology and Pharmacology, Universite Cheikh Anta Diop, Dakar, Senegal., Siddle KJ; Infectious Disease and Microbiome Program, Eli and Edythe L. Broad Institute of Harvard and MIT, Cambridge, Massachusetts, USA.; Center for Systems Biology, Department of Organismic and Evolutionary Biology, Harvard University, Cambridge, Massachusetts, USA., Mehta SB; Infectious Disease and Microbiome Program, Eli and Edythe L. Broad Institute of Harvard and MIT, Cambridge, Massachusetts, USA.; Division of Infectious Diseases, Beth Israel Deaconess Medical Center, Boston, Massachusetts, USA., Harris JB; Department of Pediatrics, Massachusetts General Hospital for Children, Boston, Massachusetts, USA., Ryan ET; Department of Medicine, Division of Infectious Diseases, Massachusetts General Hospital, Boston, Massachusetts, USA., Pierce VM; Department of Pathology and Harvard Medical School, Massachusetts General Hospital, Boston, Massachusetts, USA.; Department of Pediatrics, Massachusetts General Hospital for Children, Boston, Massachusetts, USA., LaRocque RC; Department of Medicine, Division of Infectious Diseases, Massachusetts General Hospital, Boston, Massachusetts, USA., Lemieux JE; Infectious Disease and Microbiome Program, Eli and Edythe L. Broad Institute of Harvard and MIT, Cambridge, Massachusetts, USA pardis@broadinstitute.org lemieux@broadinstitute.org Branda.John@mgh.harvard.edu Turbett.Sarah@mgh.harvard.edu.; Department of Medicine, Division of Infectious Diseases, Massachusetts General Hospital, Boston, Massachusetts, USA., Sabeti PC; Infectious Disease and Microbiome Program, Eli and Edythe L. Broad Institute of Harvard and MIT, Cambridge, Massachusetts, USA pardis@broadinstitute.org lemieux@broadinstitute.org Branda.John@mgh.harvard.edu Turbett.Sarah@mgh.harvard.edu.; Center for Systems Biology, Department of Organismic and Evolutionary Biology, Harvard University, Cambridge, Massachusetts, USA.; Immunology and Infectious Disease, Harvard T.H. Chan School of Public Health, Boston, Massachusetts, USA.; Howard Hughes Medical Institute, Chevy Chase, Maryland, USA., Rosenberg ES; Department of Pathology and Harvard Medical School, Massachusetts General Hospital, Boston, Massachusetts, USA.; Department of Medicine, Division of Infectious Diseases, Massachusetts General Hospital, Boston, Massachusetts, USA., Branda JA; Department of Pathology and Harvard Medical School, Massachusetts General Hospital, Boston, Massachusetts, USA pardis@broadinstitute.org lemieux@broadinstitute.org Branda.John@mgh.harvard.edu Turbett.Sarah@mgh.harvard.edu., Turbett SE; Department of Pathology and Harvard Medical School, Massachusetts General Hospital, Boston, Massachusetts, USA pardis@broadinstitute.org lemieux@broadinstitute.org Branda.John@mgh.harvard.edu Turbett.Sarah@mgh.harvard.edu.; Department of Medicine, Division of Infectious Diseases, Massachusetts General Hospital, Boston, Massachusetts, USA.
Jazyk:	angličtina
Zdroj:	Journal of clinical pathology [J Clin Pathol] 2021 Aug; Vol. 74 (8), pp. 496-503. Date of Electronic Publication: 2021 May 28.
DOI:	10.1136/jclinpath-2020-207128
Abstrakt:	Developing and deploying new diagnostic tests are difficult, but the need to do so in response to a rapidly emerging pandemic such as COVID-19 is crucially important. During a pandemic, laboratories play a key role in helping healthcare providers and public health authorities detect active infection, a task most commonly achieved using nucleic acid-based assays. While the landscape of diagnostics is rapidly evolving, PCR remains the gold-standard of nucleic acid-based diagnostic assays, in part due to its reliability, flexibility and wide deployment. To address a critical local shortage of testing capacity persisting during the COVID-19 outbreak, our hospital set up a molecular-based laboratory developed test (LDT) to accurately and safely diagnose SARS-CoV-2. We describe here the process of developing an emergency-use LDT, in the hope that our experience will be useful to other laboratories in future outbreaks and will help to lower barriers to establishing fast and accurate diagnostic testing in crisis conditions. Competing Interests: Competing interests: PCS is a founder and shareholder of Sherlock Biosciences, and is both on the Board and serves as shareholder of the Danaher Corporation. JEL is a consultant for Sherlock Biosciences. MNA is a cofounder, equity holder and consultant for Day Zero Diagnostics. PCS, ETR and SET have received CDC funding for this work and other COVID-related work. John Branda has received grant support from Zeus Scientific, bioMerieux, Immunetics, the Bay Area Lyme Foundation and the Lyme Disease Biobank Foundation for work unrelated to this study, and has been a consultant for T2 Biosystems, DiaSorin and Roche Diagnostics. (© Author(s) (or their employer(s)) 2021. Re-use permitted under CC BY-NC. No commercial re-use. See rights and permissions. Published by BMJ.)
Databáze:	MEDLINE
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