tetano
Editor, Senior Moderator
Chem Eng Sci
. 2022 Jan 13;117430.
doi: 10.1016/j.ces.2022.117430. Online ahead of print.
Optimization of loop-mediated isothermal amplification (LAMP) assay for robust visualization in SARS-CoV-2 and emerging variants diagnosis
Zhen Luo[SUP] 1 2 [/SUP], Chunhong Ye[SUP] 1 [/SUP], Heng Xiao[SUP] 1 2 [/SUP], Jialing Yin[SUP] 1 [/SUP], Yicong Liang[SUP] 1 [/SUP], Zhihui Ruan[SUP] 2 [/SUP], Danju Luo[SUP] 3 [/SUP], Daolong Gao[SUP] 4 [/SUP], Qiuping Tan[SUP] 4 [/SUP], Yongkui Li[SUP] 1 2 [/SUP], Qiwei Zhang[SUP] 1 2 [/SUP], Weiyong Liu[SUP] 5 [/SUP], Jianguo Wu[SUP] 1 2 4 [/SUP]
Affiliations
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 locating 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 a promising test in COVID-19 surveillance.
Keywords: Coronavirus disease 2019 (COVID-19) pandemic; Loop-mediated isothermal amplification (LAMP); Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) diagnosis; emerging SARS-CoV-2 variants.
. 2022 Jan 13;117430.
doi: 10.1016/j.ces.2022.117430. Online ahead of print.
Optimization of loop-mediated isothermal amplification (LAMP) assay for robust visualization in SARS-CoV-2 and emerging variants diagnosis
Zhen Luo[SUP] 1 2 [/SUP], Chunhong Ye[SUP] 1 [/SUP], Heng Xiao[SUP] 1 2 [/SUP], Jialing Yin[SUP] 1 [/SUP], Yicong Liang[SUP] 1 [/SUP], Zhihui Ruan[SUP] 2 [/SUP], Danju Luo[SUP] 3 [/SUP], Daolong Gao[SUP] 4 [/SUP], Qiuping Tan[SUP] 4 [/SUP], Yongkui Li[SUP] 1 2 [/SUP], Qiwei Zhang[SUP] 1 2 [/SUP], Weiyong Liu[SUP] 5 [/SUP], Jianguo Wu[SUP] 1 2 4 [/SUP]
Affiliations
- PMID: 35043022
- PMCID: PMC8757654
- DOI: 10.1016/j.ces.2022.117430
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 locating 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 a promising test in COVID-19 surveillance.
Keywords: Coronavirus disease 2019 (COVID-19) pandemic; Loop-mediated isothermal amplification (LAMP); Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) diagnosis; emerging SARS-CoV-2 variants.