The global outbreak of coronavirus disease 19(COVID-19),caused by severe acute respiratory syndrome coronavirus 2(SARS-Co V-2),has raised significant global apprehension.Developing a rapid,efficient,sensitive,and accu...The global outbreak of coronavirus disease 19(COVID-19),caused by severe acute respiratory syndrome coronavirus 2(SARS-Co V-2),has raised significant global apprehension.Developing a rapid,efficient,sensitive,and accurate point-of-care detection method is imperative for curbing SARS-Co V-2 transmission.Here,we screened a sequence,designed a set of highly sensitive loopmediated isothermal amplification primers(LAMP)and g RNA,and developed a user-friendly detection platform combining CRISPRCas12a and RT-LAMP technology to specifically detect SARS-Co V-2 and its 5 variants.Bioinformatics analysis and Cas12a-g RNA identification ensured sequence specificity,allowing us to identify SARS-Co V-2 mutations.We developed a method for the detection of SARSCoV-2 using these primers in combination with LAMP amplification and CRISPR-Cas12a technology.This method is designed to detect SARS-CoV-2(NC_045512),Alpha(B.1.1.7),Beta(B.1.351),Gamma(P.1),Delta(B.1.617.2)and Omicron(B.1.1.529).Additionally,it can differentiate SARS-CoV-2 from other coronaviruses.Quantitative analysis can be conducted by measuring fluorescence values,while qualitative analysis can be performed by observing fluorescence color point-of-care diagnosis changes with the naked eye.These results suggest that a set of novel sensitive LAMP primers and g RNA have been obtained to detect the extensive variants,and the RT-LAMPCRISPR-Cas12a platform significantly facilitates point-of-care diagnosis,thereby halting the spread of SARS-Co V-2,thus contributing to COVID-19 prevention and control.展开更多
基金Supported by the National Natural Sciences Foundation of China(52073022)the Fundamental Research Funds for the Central Universities of China and the Translational Medical Research Fund of Wuhan University Taikang Medical School(School of Basic Medical Sciences)the Key Laboratory of Environmental Pollution Monitoring and Disease Control(Guizhou Medical University)Ministry of Education(GMU-2022-HJZ)。
文摘The global outbreak of coronavirus disease 19(COVID-19),caused by severe acute respiratory syndrome coronavirus 2(SARS-Co V-2),has raised significant global apprehension.Developing a rapid,efficient,sensitive,and accurate point-of-care detection method is imperative for curbing SARS-Co V-2 transmission.Here,we screened a sequence,designed a set of highly sensitive loopmediated isothermal amplification primers(LAMP)and g RNA,and developed a user-friendly detection platform combining CRISPRCas12a and RT-LAMP technology to specifically detect SARS-Co V-2 and its 5 variants.Bioinformatics analysis and Cas12a-g RNA identification ensured sequence specificity,allowing us to identify SARS-Co V-2 mutations.We developed a method for the detection of SARSCoV-2 using these primers in combination with LAMP amplification and CRISPR-Cas12a technology.This method is designed to detect SARS-CoV-2(NC_045512),Alpha(B.1.1.7),Beta(B.1.351),Gamma(P.1),Delta(B.1.617.2)and Omicron(B.1.1.529).Additionally,it can differentiate SARS-CoV-2 from other coronaviruses.Quantitative analysis can be conducted by measuring fluorescence values,while qualitative analysis can be performed by observing fluorescence color point-of-care diagnosis changes with the naked eye.These results suggest that a set of novel sensitive LAMP primers and g RNA have been obtained to detect the extensive variants,and the RT-LAMPCRISPR-Cas12a platform significantly facilitates point-of-care diagnosis,thereby halting the spread of SARS-Co V-2,thus contributing to COVID-19 prevention and control.
