Optimization and clinical validation of dual-target RT-LAMP for SARS-CoV-2

Autor: Thomas P. Griener, Kanti Pabbaraju, Lisa Oberding, Markus Czub, Dylan R. Pillai, Guido van Marle, Keith R. Jerome, Venice Servellita, Cody Doolan, Byron M. Berenger, Luiz F. Lisboa, Charles Y. Chiu, Alexander L. Greninger, Jana Hundt, Abu Naser Mohon
Jazyk: angličtina
Rok vydání: 2020
Předmět:
0301 basic medicine
Point-of-Care Systems
Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2)
Pneumonia
Viral
030106 microbiology
education
Loop-mediated isothermal amplification
Biology
Sensitivity and Specificity
Article
Betacoronavirus
Coronavirus Envelope Proteins
03 medical and health sciences
chemistry.chemical_compound
COVID-19 Testing
Viral Envelope Proteins
LAMP
Virology
RNA polymerase
Validation
Humans
Computer Simulation
molecular
Pandemics
Gene
Diagnostics
Point of care
Detection limit
Clinical Laboratory Techniques
SARS-CoV-2
RNA-Directed DNA Polymerase
COVID-19
covid
Nucleic acid amplification technique
assay
Molecular biology
3. Good health
030104 developmental biology
PCR
Molecular Diagnostic Techniques
chemistry
Spike Glycoprotein
Coronavirus
RNA
Viral
Coronavirus Infections
Nucleic Acid Amplification Techniques
Zdroj: Journal of Virological Methods
ISSN: 1879-0984
0166-0934
Popis: Highlights • The LAMP method performs equally to international reference RT-PCR methods for detection of SARS-CoV-2 • Experiments have been conducted that fulfil regulatory criteria by international bodies • LAMP as an LDT does not rely on RT-PCR reagents and does not cannibalize key reagents and kits in those supply chains • The test is rapid with a result in less than 30 minutes after extraction • Many countries can use this LDT option internationally in one or other format and provide vital testing • Potential to apply this chemistry to a point of care method exists
A novel reverse-transcriptase loop mediated amplification (RT-LAMP) method targeting genes encoding the Spike (S) protein and RNA-dependent RNA polymerase (RdRP) of SARS-CoV-2 has been developed. The LAMP assay achieves a comparable limit of detection (25 copies per reaction) as commonly used RT-PCR protocols using clinical samples quantified by digital droplet PCR. Precision, cross-reactivity, inclusivity, and limit of detection studies were performed according to regulatory standards. Clinical validation of dual-target RT-LAMP (S and RdRP gene) achieved a PPA of 98.48% (95% CI 91.84% to 99.96%) and NPA 100.00% (95% CI 93.84% to 100.00%) based on the E gene and N2 gene reference RT-PCR methods. The method has implications for development of point of care technology using isothermal amplification.
Databáze: OpenAIRE
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