RPB0037

Pathogen Description

Target Pathogen Pathogen Name NCBI Taxonomy ID Order Family Genus Species Pathogen type
SARS-CoV-2 SARS-CoV-2, 2019-nCoV, COVID-19, COVID-19 virus, SARS2, Wuhan coronavirus, Human coronavirus 2019, COVID19, HCoV-19, SARS-2, SARS-CoV4 2697049 Nidovirales Coronaviridae Betacoronavirus Severe acute respiratory syndrome-related coronavirus virus

Primer Description

Primer Name Sequence(5'-3') Length(bp) Primer Final Concentration(μM） GC Content（%） Predicted Melting Temperature(℃） Molecular Weight(g/moles) Positions in GenBank accession number
F GAAATATACTACGACGACTGATCACATACTACTTCTGTTTACATAGA 47 0.16 34 60 14388.44 \
R TCCTAGGTTGAAGATAACCCACATAATAAG 30 0.16 36.7 55.18 9207.08 \

Gene Description

Target Gene GenBank ID
S \

Assay Description

Application Assay Primer Designing Software Reaction Time(min) Assay Temperature(℃) Readout System(s) Limit of Detection(LoD) Sensitivity(%) Specificity(%)
sensitively deteceing SARS-CoV-2 CRISPR/Cas13a-RPA \ 20 37 fluorescence intensity of the biosensor/LF APHFanalyzer:0.68 fM;lateral flow strip:2 fM \ \

Publication Description

Year of Publication Title Author(s) Journal PMID DOI
2022 Automated, portable, and high-throughput fluorescence analyzer (APHF-analyzer) and lateral flow strip based on CRISPR/Cas13a for sensitive and visual detection of SARS-CoV-2 Gaihua Cao, Danqun Huo, Xiaolong Chen, Xianfeng Wang, Shiying Zhou, Shixian Zhao, Xiaogang Luo, Changjun Hou Talanta 35653961 10.1016/j.talanta.2022.123594

Automated, portable, and high-throughput fluorescence analyzer (APHF-analyzer) and lateral flow strip based on CRISPR/Cas13a for sensitive and visual detection of SARS-CoV-2

Author(s):

Gaihua Cao, Danqun Huo, Xiaolong Chen, Xianfeng Wang, Shiying Zhou, Shixian Zhao, Xiaogang Luo, Changjun Hou

Journal:

Talanta

Year:

2022

Abstract:

COVID-19 has erupted and quickly swept across the globe, causing huge losses to human health and wealth. It is of great value to develop a quick, accurate, visual, and high-throughput detection of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). Here, we developed a biosensor based on CRISPR/Cas13a combined with recombinase polymerase amplification (RPA) to detect S and Orf1ab genes of SARS-CoV-2 within 30 min. Most important of all, we developed an automated, portable, and high-throughput fluorescence analyzer (APHF-analyzer) with a 3D-printed microfluidic chip for sensitively detecting SARS-CoV-2, which addressed aerosol contamination issue and provided a more accurate and high-throughput detection during the on-site detection process. The detection limits of S gene and Orf1ab gene were as low as 0.68 fM and 4.16 fM. Furthermore, we used the lateral flow strip to realize visualization and point of care testing (POCT) of SARS-CoV-2. Therefore, profit from the efficient amplification of RPA and the high specificity of CRISPR/Cas13a, APHF-analyzer and the lateral flow strip to simultaneous detection of S gene and Orf1ab gene would be applied as a promising tool in the field of SARS-CoV-2 detection.