tetano
Editor, Senior Moderator
Biotechniques
. 2022 May 12.
doi: 10.2144/btn-2022-0037. Online ahead of print.
Isothermal amplification using sequence-specific fluorescence detection of SARS coronavirus 2 and variants in nasal swabs
Les Jones[SUP] 1 [/SUP], Hemant K Naikare[SUP] 2 [/SUP], Yung-Yi C Mosley[SUP] 2 [/SUP], Ralph A Tripp[SUP] 1 [/SUP]
Affiliations
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.
Keywords: FQ-LAMP; POC; RT-LAMP; SARS coronavirus 2; diagnostics; fluorescence; variant.
. 2022 May 12.
doi: 10.2144/btn-2022-0037. Online ahead of print.
Isothermal amplification using sequence-specific fluorescence detection of SARS coronavirus 2 and variants in nasal swabs
Les Jones[SUP] 1 [/SUP], Hemant K Naikare[SUP] 2 [/SUP], Yung-Yi C Mosley[SUP] 2 [/SUP], Ralph A Tripp[SUP] 1 [/SUP]
Affiliations
- PMID: 35545967
- DOI: 10.2144/btn-2022-0037
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.
Keywords: FQ-LAMP; POC; RT-LAMP; SARS coronavirus 2; diagnostics; fluorescence; variant.