학술논문

SARS-CoV-2 viral variants can rapidly be identified for clinical decision making and population surveillance using a high-throughput digital droplet PCR assay
Document Type
article
Source
Scientific Reports. 13(1)
Subject
Microbiology
Biological Sciences
Biomedical and Clinical Sciences
Prevention
Genetics
Biodefense
Vaccine Related
Biotechnology
Emerging Infectious Diseases
Infection
Good Health and Well Being
Humans
SARS-CoV-2
COVID-19
Polymerase Chain Reaction
Clinical Decision-Making
Population Surveillance
COVID-19 Testing
U. S. C. Variant Study Group
Language
Abstract
Epidemiologic surveillance of circulating SARS-CoV-2 variants is essential to assess impact on clinical outcomes and vaccine efficacy. Whole genome sequencing (WGS), the gold-standard to identify variants, requires significant infrastructure and expertise. We developed a digital droplet polymerase chain reaction (ddPCR) assay that can rapidly identify circulating variants of concern/interest (VOC/VOI) using variant-specific mutation combinations in the Spike gene. To validate the assay, 800 saliva samples known to be SARS-CoV-2 positive by RT-PCR were used. During the study (July 2020-March 2022) the assay was easily adaptable to identify not only existing circulating VAC/VOI, but all new variants as they evolved. The assay can discriminate nine variants (Alpha, Beta, Gamma, Delta, Eta, Epsilon, Lambda, Mu, and Omicron) and sub-lineages (Delta 417N, Omicron BA.1, BA.2). Sequence analyses confirmed variant type for 124/124 samples tested. This ddPCR assay is an inexpensive, sensitive, high-throughput assay that can easily be adapted as new variants are identified.