RPB0033

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 TACACCGGAAGCCAATATGGATCAAGAATC 30 0.4 43.3 59.89 9217.08 \
R CATTAGCACAAGTTGTAGGTATTTGTACATAC 32 0.4 34.4 54.99 9837.47 \

Gene Description

Target Gene GenBank ID
ORF1ab \

Assay Description

Application Assay Primer Designing Software Reaction Time(min) Assay Temperature(℃) Readout System(s) Limit of Detection(LoD) Sensitivity(%) Specificity(%)
sensitive COVID-19 diagnosis RT-RPA-OC-MLFA \ 20 37 the line intensity was photographed and analyzed with Image J 10 copies per reaction (30 μL) \ \

Publication Description

Year of Publication Title Author(s) Journal PMID DOI
2022 Multiplexed lateral flow assay integrated with orthogonal CRISPR-Cas system for SARS-CoV-2 detection Gaoxing Su,Min Zhu,Diyuan Li,Mengting Xu,Yuedong Zhu,Yan Zhang,Hongyan Zhu,Feng Li,Yanyan Yu Sensors and Actuators:B. Chemical 36032355 10.1016/j.snb.2022.132537

Multiplexed lateral flow assay integrated with orthogonal CRISPR-Cas system for SARS-CoV-2 detection

Author(s):

Gaoxing Su,Min Zhu,Diyuan Li,Mengting Xu,Yuedong Zhu,Yan Zhang,Hongyan Zhu,Feng Li,Yanyan Yu

Journal:

Sensors and Actuators:B. Chemical

Year:

2022

Abstract:

The development of field-deployable detection platform amenable for multiplexed genes testing will significantly improve the efficiency and reliability during point-of-care testing (POCT) applications. In this regard, an orthogonal CRISPR-Cas-mediated multiplexed lateral flow assay (designated as OC-MLFA) is proposed for SARS-CoV-2 genome detection. Taking the advantage of activation and cleavage preferences between Cas12a and Cas13a, orthogonal (two-independent-channel signal readout) CRISPR-Cas system is investigated. Lateral flow strips with two target lines are designed to accommodate the orthogonal CRISPR system. The interference between Cas12a and Cas13a channels can be effectively eliminated via the elaborate nucleic acids and lateral flow strips design. The high preamplification efficiency from reverse transcription recombinase polymerase amplification (RT-RPA) and Cas enzyme mediated trans-cleavage process bring the sensitivity of our OC-MLFA method to 10 copies per test (30 μL). Nasopharyngeal swab clinical samples with different cycle threshold (Ct) values according to the RT-PCR method were analyzed with the proposed OC-MLFA, during which 76 out of 76 detection accuracy was obtained. Featured with the multiplexed genes detection simultaneously in one reaction and colorimetric readout through single strip, the OC-MLFA we proposed herein ensures great accuracy and efficiency, which endows promising field-deployable POCT application feasibility.