Optimization of loop-mediated isothermal amplification (LAMP) assay for robust visualization in SARS-CoV-2 and emerging variants diagnosis.
Chem Eng Sci
; 251: 117430, 2022 Apr 06.
Article
em En
| MEDLINE
| ID: mdl-35043022
Loop-mediated isothermal amplification (LAMP) is widely used in detection of pathogenic microorganisms including SARS-CoV-2. However, the performance of LAMP assay needs further exploration in the emerging SARS-CoV-2 variants test. Here, we design serials of primers and select an optimal set for LAMP-based on SARS-CoV-2 N gene for a robust and visual assay in SARS-CoV-2 diagnosis. The limit of detectable template reaches 10 copies of N gene per 25 µL reaction at isothermal 58â within 40 min. Importantly, the primers for LAMP assay locate at 12 to 213 nt of N gene, a highly conservative region, which serves as a compatible test in emerging SARS-CoV-2 variants. Comparison to a commercial qPCR assay, this LAMP assay exerts the high viability in diagnosis of 41 clinical samples. Our study optimizes an advantageous LAMP assay for colorimetric detection of SARS-CoV-2 and emerging variants, which is hopeful to be a promising test in COVID-19 surveillance.
COVID-19, coronavirus disease 2019; CRISPR, clustered regularly interspaced short palindromic repeats; Coronavirus disease 2019 (COVID-19) pandemic; Ct, threshold cycle; Emerging SARS-CoV-2 variants; IVD, in-vitro diagnosis; LAMP, Loop-mediated isothermal amplification; Loop-mediated isothermal amplification (LAMP); NGS, next-generation sequencing; POC, point-of-care; RT-qPCR, real-time reverse transcriptase quantitative polymerase chain reaction; SARS-CoV-2, severe acute respiratory syndrome coronavirus 2; Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) diagnosis; VOC, variants of concern
Texto completo:
1
Coleções:
01-internacional
Base de dados:
MEDLINE
Tipo de estudo:
Diagnostic_studies
Idioma:
En
Ano de publicação:
2022
Tipo de documento:
Article