RPB0015

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 CCTACAACTTGTGCTAATGACCCTGTGGGTTTTACACTT 39 \ 43.6 64.69 11898.77 \
R CACCGCAAACCCGTTTAAAAACGATTGTGC 30 \ 46.7 62.91 9144.02 \
P ACACAGTCTGTACCGTCTGCGGTATGTGGAAAGG[HEX-dT][THF]A[BHQ1-dT]GGCTGTAGTTGTGATCAAC[phosphate] 54 \ 48.2 71.17 17365.29 \

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(%)
significantly contribute to detecting patients with COVID-19 in low-resource settings and point-of-care testing (POCT) centrifugal microfluidics-based multiplexRT-RPA \ 20 \ real time fluorescence 10 RNA copies/reaction 95 100

Publication Description

Year of Publication Title Author(s) Journal PMID DOI
2023 Centrifugal microfluidic-based multiplex recombinase polymerase amplification assay for rapid detection of SARS-CoV-2 Ruoxu Li,Ning Su,Xiaodong Ren,Xiange Sun,Wenman Li,Yuwei Li,Jin Li,Chen Chen,Hong Wang,Weiping Lu,Shaoli Deng,Qing Huang iScience 36845031 10.1016/j.isci.2023.106245

Centrifugal microfluidic-based multiplex recombinase polymerase amplification assay for rapid detection of SARS-CoV-2

Author(s):

Ruoxu Li,Ning Su,Xiaodong Ren,Xiange Sun,Wenman Li,Yuwei Li,Jin Li,Chen Chen,Hong Wang,Weiping Lu,Shaoli Deng,Qing Huang

Journal:

iScience

Year:

2023

Abstract:

The COVID-19 pandemic has spread worldwide, and rapid detection of the SARS-CoV-2 virus is crucial for infection surveillance and epidemic control. This study developed a centrifugal microfluidics-based multiplex reverse transcription recombinase polymerase amplification (RT-RPA) assay for endpoint fluorescence detection of the E, N, and ORF1ab genes of SARS-CoV-2. The microscope slide-shaped microfluidic chip could simultaneously accomplish three target genes and one reference human gene (i.e., ACTB) RT-RPA reactions in 30 min, and the sensitivity was 40 RNA copies/reaction for the E gene, 20 RNA copies/reaction for the N gene, and 10 RNA copies/reaction for the ORF1ab gene. The chip demonstrated high specificity, reproducibility, and repeatability. Chip performance was also evaluated using real clinical samples. Thus, this rapid, accurate, on-site, and multiplexed nucleic acid test microfluidic chip would significantly contribute to detecting patients with COVID-19 in low-resource settings and point-of-care testing (POCT) and, in the future, could be used to detect emerging new variants of SARS-CoV-2.