期刊
BIOSENSORS & BIOELECTRONICS
卷 198, 期 -, 页码 -出版社
ELSEVIER ADVANCED TECHNOLOGY
DOI: 10.1016/j.bios.2021.113857
关键词
CRISPR/Cas12; RT-CORDS; D614G; N501Y; 69/70 deletion; SARS-CoV-2 variants
类别
资金
- National Natural Science Foundation of China [31871292]
- Fundamental Research Funds for the Central Universities [2019MS089]
- South China University of Technology [2019A1515010619]
- Natural Science Foundation of Guangdong Province of China [A2019432]
- Medical Scien-tific Research Foundation of Guangdong Province of China [2018YFC0910201]
- National Key R&D Program of China
The increasing prevalence of SARS-CoV-2 variants with spike mutations has raised concerns. Rapid and accurate detection of these variants is crucial for infection prevention and control. The RT-CORDS platform accurately, sensitively, specifically, and rapidly detects SARS-CoV-2 variants, making it a potential tool for clinical diagnosis.
The increasing prevalence of SARS-CoV-2 variants with spike mutations has raised concerns owing to higher transmission rates, disease severity, and escape from neutralizing antibodies. Rapid and accurate detection of SARS-CoV-2 variants provides crucial information concerning the outbreaks of SARS-CoV-2 variants and possible lines of transmission. This information is vital for infection prevention and control. We used a Cas12a-based RT-PCR combined with CRISPR on-site rapid detection system (RT-CORDS) platform to detect the key mutations in SARS-CoV-2 variants, such as 69/70 deletion, N501Y, and D614G. We used type-specific CRISPR RNAs (crRNAs) to identify wild-type (crRNA-W) and mutant (crRNA-M) sequences of SARS-CoV-2. We successfully differentiated mutant variants from wild-type SARS-CoV-2 with a sensitivity of 10(-17) M (approximately 6 copies/mu L). The assay took just 10 min with the Cas12a/crRNA reaction after a simple RT-PCR using a fluorescence reporting system. In addition, a sensitivity of 10(-16) M could be achieved when lateral flow strips were used as readouts. The accuracy of RT-CORDS for SARS-CoV-2 variant detection was 100% consistent with the sequencing data. In conclusion, using the RT-CORDS platform, we accurately, sensitively, specifically, and rapidly detected SARS-CoV-2 variants. This method may be used in clinical diagnosis.
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