Isothermal amplification using sequence-specific fluorescence detection of SARS coronavirus 2 and variants in nasal swabs

Author:

Jones Les1,Naikare Hemant K2,Mosley Yung-Yi C2,Tripp Ralph A1ORCID

Affiliation:

1. Department of Infectious Disease, College of Veterinary Medicine, University of Georgia, Athens, GA, USA

2. Tifton Veterinary Diagnostic and Investigational Laboratory, University of Georgia, Athens, GA, USA

Abstract

Coronavirus disease 2019 is a public health challenge requiring rapid testing for the detection of infections and transmission. Nucleic acid amplification tests targeting SARS coronavirus 2 (CoV2) are used to detect CoV2 in clinical samples. Real-time reverse transcription quantitative PCR is the standard nucleic acid amplification test for CoV2, although reverse transcription loop-mediated isothermal amplification is used in diagnostics. The authors demonstrate a sequence-specific reverse transcription loop-mediated isothermal amplification-based nucleic acid amplification assay that is finished within 30 min using minimally processed clinical nasal swab samples and describe a fluorescence-quenched reverse transcription loop-mediated isothermal amplification assay using labeled primers and a quencher oligonucleotide. This assay can achieve rapid (30 min) and sensitive (1000 plaque-forming units/ml) fluorescence detection of CoV2 (WA1/2020), B.1.1.7 (Alpha) and variants of concern Delta (B.1.617.2) and Omicron (B.1.1.529) in nasal samples.

Publisher

Future Science Ltd

Subject

General Biochemistry, Genetics and Molecular Biology,Biotechnology

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