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 assay.

Autor: Anahtar MN; Department of Pathology and Harvard Medical School, Massachusetts General Hospital, Boston, Massachusetts., Shaw B; Broad Institute of Massachusetts Institute of Technology (MIT) and Harvard, Cambridge, MA, USA.; Department of Medicine, Division of Infectious Diseases, Massachusetts General Hospital, Boston, MA, USA., Slater D; Department of Medicine, Division of Infectious Diseases, Massachusetts General Hospital, Boston, MA, USA., Byrne E; Broad Institute of Massachusetts Institute of Technology (MIT) and Harvard, Cambridge, MA, USA., Botti-Lodovico Y; Broad Institute of Massachusetts Institute of Technology (MIT) and Harvard, Cambridge, MA, USA., Adams G; Broad Institute of Massachusetts Institute of Technology (MIT) and Harvard, Cambridge, MA, USA.; Department of Medicine, Division of Infectious Diseases, Massachusetts General Hospital, Boston, MA, USA., Schaffner S; Broad Institute of Massachusetts Institute of Technology (MIT) and Harvard, Cambridge, MA, USA.; Center for Systems Biology, Department of Organismic and Evolutionary Biology, Harvard University, Cambridge, Massachusetts, USA.; Department of Immunology and Infectious Diseases, Harvard T.H. Chan School of Public Health, Harvard University, Boston, Massachusetts, USA., Eversley J; Department of Pathology and Harvard Medical School, Massachusetts General Hospital, Boston, Massachusetts., McGrath G; Department of Medicine, Division of Infectious Diseases, Massachusetts General Hospital, Boston, MA, USA., Gogakos T; Department of Pathology and Harvard Medical School, Massachusetts General Hospital, Boston, Massachusetts., Lennerz J; Department of Pathology and Harvard Medical School, Massachusetts General Hospital, Boston, Massachusetts., Desai Marble H; Department of Pathology and Harvard Medical School, Massachusetts General Hospital, Boston, Massachusetts., Ritterhouse LL; Department of Pathology and Harvard Medical School, Massachusetts General Hospital, Boston, Massachusetts., Batten J; Department of Pathology and Harvard Medical School, Massachusetts General Hospital, Boston, Massachusetts., Georgantas NZ; Department of Pathology and Harvard Medical School, Massachusetts General Hospital, Boston, Massachusetts., Pellerin R; Department of Pathology and Harvard Medical School, Massachusetts General Hospital, Boston, Massachusetts., Signorelli S; Department of Pathology and Harvard Medical School, Massachusetts General Hospital, Boston, Massachusetts., Thierauf J; Department of Pathology and Harvard Medical School, Massachusetts General Hospital, Boston, Massachusetts.; Department of Otorhinolaryngology, Head and Neck Surgery, Experimental Head and Neck Oncology, Heidelberg University Hospital, Heidelberg, Germany., Kemball M; Broad Institute of Massachusetts Institute of Technology (MIT) and Harvard, Cambridge, MA, 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 State, Nigeria.; African Center of Excellence for Genomics of Infectious Diseases (ACEGID), Redeemer's University, Ede, Osun State, Nigeria., Grant DS; Viral Hemorrhagic Fever Program, Kenema Government Hospital, Ministry of Health and Sanitation, 1 Combema Road, 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 (ACEGID), Redeemer's University, Ede, Osun State, Nigeria.; Université Cheikh Anta Diop, BP 5005, Dakar, Sénégal., Siddle KJ; Broad Institute of Massachusetts Institute of Technology (MIT) and Harvard, Cambridge, MA, USA.; Center for Systems Biology, Department of Organismic and Evolutionary Biology, Harvard University, Cambridge, Massachusetts, USA., Mehta SB; Broad Institute of Massachusetts Institute of Technology (MIT) and Harvard, Cambridge, MA, USA.; Division of Infectious Diseases, Beth Israel Deaconess Medical Center, Boston, MA, USA., Harris J; Department of Pediatrics, Massachusetts General Hospital for Children, Boston, MA, USA., Ryan ET; Department of Medicine, Division of Infectious Diseases, Massachusetts General Hospital, Boston, MA, USA., Pierce V; Department of Pathology and Harvard Medical School, Massachusetts General Hospital, Boston, Massachusetts.; Department of Pediatrics, Massachusetts General Hospital for Children, Boston, MA, USA., LaRocque R; Department of Medicine, Division of Infectious Diseases, Massachusetts General Hospital, Boston, MA, USA., Lemieux JE; Broad Institute of Massachusetts Institute of Technology (MIT) and Harvard, Cambridge, MA, USA.; Department of Medicine, Division of Infectious Diseases, Massachusetts General Hospital, Boston, MA, USA., Sabeti P; Broad Institute of Massachusetts Institute of Technology (MIT) and Harvard, Cambridge, MA, USA.; Department of Medicine, Division of Infectious Diseases, Massachusetts General Hospital, Boston, MA, USA.; Center for Systems Biology, Department of Organismic and Evolutionary Biology, Harvard University, Cambridge, Massachusetts, USA.; Department of Immunology and Infectious Diseases, Harvard T.H. Chan School of Public Health, Harvard University, Boston, Massachusetts, USA.; Howard Hughes Medical Institute, Chevy Chase, Maryland.; Massachusetts Consortium on Pathogen Readiness, Boston, MA, USA., Rosenberg E; Department of Pathology and Harvard Medical School, Massachusetts General Hospital, Boston, Massachusetts.; Department of Medicine, Division of Infectious Diseases, Massachusetts General Hospital, Boston, MA, USA., Branda J; Department of Pathology and Harvard Medical School, Massachusetts General Hospital, Boston, Massachusetts., Turbett SE; Department of Pathology and Harvard Medical School, Massachusetts General Hospital, Boston, Massachusetts.; Department of Medicine, Division of Infectious Diseases, Massachusetts General Hospital, Boston, MA, USA.
Jazyk: angličtina
Zdroj: MedRxiv : the preprint server for health sciences [medRxiv] 2020 Sep 01. Date of Electronic Publication: 2020 Sep 01.
DOI: 10.1101/2020.08.26.20157297
Abstrakt: Developing and deploying new diagnostic tests is difficult, but the need to do so in response to a rapidly emerging pandemic such as COVID-19 is crucially important for an effective response. In the early stages of a pandemic, laboratories play a key role in helping health care 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, polymerase chain reaction (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 fast and accurate diagnostic testing in crisis conditions.
Databáze: MEDLINE