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Optimization of loop-mediated isothermal amplification (LAMP) assay for robust visualization in SARS-CoV-2 and emerging variants diagnosis

Chem Eng Sci. 2022-01; 
Zhen Luo, Chunhong Ye, Heng Xiao, Jialing Yin, Yicong Liang, Zhihui Ruan, Danju Luo, Daolong Gao, Qiuping Tan, Yongkui Li, Qiwei Zhang, Weiyong Liu, Jianguo Wu
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Gene Synthesis … NCBI GenBank database (28274 to 29533 nt in complete genome from Wuhan-Hu-1 strain, accession number: NC_045512.2.). The target gene was synthesized (GenScript Co., Ltd … Get A Quote

摘要

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

关键词

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