RPB0215

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 AGGCAGCAGTAGGGGAACTTCTCCTGCTAGAAT 33 \ 51.5 66.82 10202.68 \
R TTGGCCTTTACCAGACATTTTGCTCTCAAGCTG 33 \ 45.5 63.78 10045.57 \

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(%)
\ RT-RPA \ 15 37 Fluorometric detection (Fluorescent metal ion indicator (Calcein)) 100 copies per test \ \

Publication Description

Year of Publication Title Author(s) Journal PMID DOI
2021 Autonomous Lab-on-Paper for Multiplexed, CRISPR-based Diagnostics of SARS-CoV-2 Kun Yin , Xiong Ding , Ziyue Li , Maroun M Sfeir , Enrique Ballesteros , Changchun Liu Lab on a Chip 34100058 10.1039/d1lc00293g

Autonomous Lab-on-Paper for Multiplexed, CRISPR-based Diagnostics of SARS-CoV-2

Author(s):

Kun Yin , Xiong Ding , Ziyue Li , Maroun M Sfeir , Enrique Ballesteros , Changchun Liu

Journal:

Lab on a Chip

Year:

2021

Abstract:

The COVID-19 pandemic, caused by severe acute respiratory coronavirus 2 (SARS-CoV-2), has become a public health emergency and widely spread around the world. Rapid, accurate and early diagnosis of COVID-19 infection plays a crucial role in breaking this pandemic. However, the detection accuracy is limited for current single-gene diagnosis of SARS-CoV-2. Herein, we develop an autonomous lab-on-paper platform for multiplex gene diagnosis of SARS-CoV-2 by combining reverse transcription recombinase polymerase amplification (RT-RPA) and CRISPR-Cas12a detection. The autonomous lab-on-paper is capable of simultaneously detecting nucleoprotein (N) gene and spike (S) gene of SARS-CoV-2 virus as well as human housekeeping RNAse P gene (an internal control) in a single clinical sample. With the developed platform, 102 copies viral RNA per test can be detected within one hour. Also, the lab-on-paper platform has been used to detect 21 swab clinical samples and obtains a comparable performance to the conventional RT-PCR method. Thus, the developed lab-on-paper platform holds great potential for rapid, sensitive, reliable, multiple molecular diagnostics of COVID-19 and other infectious diseases in resource-limited settings.