RPB0367

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
SARS-CoV-2 F TGGTGGACCCTCAGATTCAACTGGCAGTAAC 31 480 nM 51.61 65.28 9520.24 \
SARS-CoV-2 R CTTGCCATGTTGAGTGAGAGCGGTGAACCAAG 32 480 nM 53.13 66.37 9929.5 \

Gene Description

Target Gene GenBank ID
N gene \

Assay Description

Application Assay Primer Designing Software Reaction Time(min) Assay Temperature(℃) Readout System(s) Limit of Detection(LoD) Sensitivity(%) Specificity(%)
Simplicity and flexibility for these three virus detections. T RPA-CRISPR-Cas12a \ 30 min 40°C CRISPR-Cas12a 10²copies\µL \ \

Publication Description

Year of Publication Title Author(s) Journal PMID DOI
2023 Development of a rapid, sensitive detection method for SARS-CoV-2 and influenza virus based on recombinase polymerase amplification combined with CRISPR-Cas12a assay Yuning Wang,Liqiang Wu,Xiaomei Yu,Gang Wang,Ting Pan,Zhao Huang,Ting Cui,Tianxun Huang,Zhentao Huang,Libo Nie,Chungen Qian Journal of Medical Virology 37933907 10.1002/jmv.29215

Development of a rapid, sensitive detection method for SARS-CoV-2 and influenza virus based on recombinase polymerase amplification combined with CRISPR-Cas12a assay

Author(s):

Yuning Wang,Liqiang Wu,Xiaomei Yu,Gang Wang,Ting Pan,Zhao Huang,Ting Cui,Tianxun Huang,Zhentao Huang,Libo Nie,Chungen Qian

Journal:

Journal of Medical Virology

Year:

2023

Abstract:

Respiratory tract infections are associated with the most common diseases transmitted among people and remain a huge threat to global public health. Rapid and sensitive diagnosis of causative agents is critical for timely treatment and disease control. Here, we developed a novel method based on recombinase polymerase amplification (RPA) combined with CRISPR-Cas12a to detect three viral pathogens, including SARS-CoV-2, influenza A, and influenza B, which cause similar symptom complexes of flu cold in the respiratory tract. The detection method can be completed within 1 h, which is faster than other standard detection methods, and the limit of detection is approximately 102 copies/μL. Additionally, this detection system is highly specific and there is no cross-reactivity with other common respiratory tract pathogens. Based on this assay, we further developed a more simplified RPA/CRISPR-Cas12a system combined with lateral flow assay on a manual microfluidic chip, which can simultaneously detect these three viruses. This low-cost detection system is rapid and sensitive, which could be applied in the field and resource-limited areas without bulky and expensive instruments, providing powerful tools for the point-of-care diagnostic.