Optimization of loop-mediated isothermal amplification (LAMP) assay for robust visualization in SARS-CoV-2 and emerging variants diagnosis
| Autor: | 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 |
|---|---|
| Rok vydání: | 2022 |
| Předmět: |
Loop-mediated isothermal amplification (LAMP)
Emerging SARS-CoV-2 variants Applied Mathematics General Chemical Engineering General Chemistry POC point-of-care Ct threshold cycle SARS-CoV-2 severe acute respiratory syndrome coronavirus 2 Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) diagnosis Article Industrial and Manufacturing Engineering NGS next-generation sequencing LAMP Loop-mediated isothermal amplification CRISPR clustered regularly interspaced short palindromic repeats RT-qPCR real-time reverse transcriptase quantitative polymerase chain reaction IVD in-vitro diagnosis VOC variants of concern ComputingMethodologies_COMPUTERGRAPHICS COVID-19 coronavirus disease 2019 Coronavirus disease 2019 (COVID-19) pandemic |
| Zdroj: | Chemical Engineering Science |
| ISSN: | 0009-2509 |
| DOI: | 10.1016/j.ces.2022.117430 |
| Popis: | Graphical abstract 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 58℃ 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. |
| Databáze: | OpenAIRE |
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