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? gene for a robust and visual assay in SARS-CoV-2 diagnosis. The limit of detectable template reaches 10 copies of gene per 25?μL reaction at isothermal 58℃ within 40?min. Importantly, the primers for LAMP assay locate at 12 to 213 nt of 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 ... More
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? gene for a robust and visual assay in SARS-CoV-2 diagnosis. The limit of detectable template reaches 10 copies of gene per 25?μL reaction at isothermal 58℃ within 40?min. Importantly, the primers for LAMP assay locate at 12 to 213 nt of 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
