Related Experiment Video
Updated: Oct 13, 2025
04:17
DNA Virus Detection System Based on RPA-CRISPR/Cas12a-SPM and Deep Learning
Published on: May 10, 2024
961
CRISPR-Cas12a-Based Detection for the Major SARS-CoV-2 Variants of Concern
Yuanhao Liang1, Hongqing Lin1, Lirong Zou2
1Department of Epidemiology, School of Public Health, Southern Medical Universitygrid.284723.8, Guangzhou, China.
Microbiology Spectrum
|November 17, 2021
Summary
A new CRISPR-Cas12a test rapidly detects SARS-CoV-2 variants by identifying key mutations. This simple assay offers 100% accuracy in clinical samples, aiding global pandemic control efforts.
Area of Science:
- Molecular Biology
- Virology
- Biotechnology
Background:
- Emergence of SARS-CoV-2 variants of concern (VOCs) poses a significant challenge to controlling the COVID-19 pandemic.
- Existing methods for variant detection can be slow and complex, creating a need for rapid and accessible diagnostic tools.
- Accurate and timely identification of SARS-CoV-2 variants is crucial for public health surveillance and response.
Purpose of the Study:
- To design and validate a CRISPR-Cas12a system for the direct and rapid detection of SARS-CoV-2 VOCs.
- To improve detection sensitivity by combining CRISPR-Cas12a with reverse transcription-PCR (RT-PCR).
- To develop a universal system capable of detecting mutations without requiring a specific protospacer adjacent motif (PAM).
Main Methods:
- Development of a CRISPR-Cas12a assay targeting specific mutations in the SARS-CoV-2 spike protein and ORF8 gene.
- Analytical validation of the CRISPR-Cas12a system for sensitivity and specificity.
- Combination of RT-PCR with CRISPR-Cas12a for enhanced detection and development of a PAM-independent detection strategy.
Main Results:
- The CRISPR-Cas12a assay successfully detected key spike protein mutations (e.g., K417N/T, L452R/Q, E484K/Q, N501Y) distinguishing multiple SARS-CoV-2 variants (Alpha, Beta, Gamma, Delta, etc.).
- The assay differentiated SARS-CoV-2 lineages based on ORF8 mutations with a low limit of detection of 10 copies/reaction.
- 100% concordance was observed between the CRISPR-based assay and sequencing when testing 32 clinical samples positive for wild-type, Alpha, Beta, and Delta variants.
Conclusions:
- The developed CRISPR-Cas12a system provides a rapid, robust, and accurate method for SARS-CoV-2 variant genotyping.
- This approach can be readily adapted for screening emerging mutations and implemented in laboratories with existing nucleic acid amplification testing capabilities.
- The system is particularly valuable for resource-limited settings, offering a scalable solution for global pandemic surveillance.
More Related Videos
10:16Author Spotlight: Development of Simplified CRISPR-Based Tests for Rapid Detection of Infectious Diseases
Published on: August 16, 2024
1.5K
07:59Rapid and Specific Detection of Acinetobacter baumannii Infections Using a Recombinase Polymerase Amplification/Cas12a-based System
Published on: April 25, 2025
646
Related Concept Videos
01:05Single Nucleotide Polymorphisms-SNPs
16.8K
A single nucleotide polymorphism or SNP is a single nucleotide variation at a specific genomic position in a large population. It is the most prevalent type of sequence variation found in the human genome. Point mutations that occur in more than 1% of the population qualify as SNPs. These are present once every 1000 nucleotides on an average in the human genome. Replacement of a purine with another purine (A/G) or a pyrimidine with another pyrimidine (C/T) is known as a transition. In contrast,...
16.8K
01:59CRISPR
53.4K
Genome editing technologies allow scientists to modify an organism’s DNA via the addition, removal, or rearrangement of genetic material at specific genomic locations. These types of techniques could potentially be used to cure genetic disorders such as hemophilia and sickle cell anemia. One popular and widely used DNA-editing research tool that could lead to safe and effective cures for genetic disorders is the CRISPR-Cas9 system. CRISPR-Cas9 stands for Clustered Regularly Interspaced...
53.4K
02:53CRISPR and crRNAs
17.8K
Bacteria and archaea are susceptible to viral infections just like eukaryotes; therefore, they have developed a unique adaptive immune system to protect themselves. Clustered regularly interspaced short palindromic repeats and CRISPR-associated proteins (CRISPR-Cas) are present in more than 45% of known bacteria and 90% of known archaea.
The CRISPR-Cas system stores a copy of foreign DNA in the host genome and uses it to identify the foreign DNA upon reinfection. CRISPR-Cas has three different...
The CRISPR-Cas system stores a copy of foreign DNA in the host genome and uses it to identify the foreign DNA upon reinfection. CRISPR-Cas has three different...
17.8K
01:28CRISPR/Cas9 Genome Editing
615
The CRISPR-Cas system serves as a bacterial defense mechanism against invading genetic elements such as viruses and plasmids, forming the foundation for its adaptation as a powerful genome-editing tool. Originally discovered in prokaryotes, this system has been repurposed to revolutionize genetic engineering across a wide range of organisms, including plants, animals, and humans. The core component, Cas9, is an endonuclease derived from Streptococcus pyogenes, capable of introducing...
615