RPB0072

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
ORF1ab-F5 TTGCCTGGCACGATATTACGCACAACTAATGGT 33 \ 45.45 64.52 10112.63 \
ORF1ab-R1 CAAGCTGATGTTGCAAAGTCAGTGTACTCTAT 32 \ 40.63 59.75 9838.46 \

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(%)
\ \ \ 20 42 CRISPR-Cas12a 1 copy/μL 100 100

Publication Description

Year of Publication Title Author(s) Journal PMID DOI
2020 Rapid detection of SARS-CoV-2 with CRISPR-Cas12a. Xiong, Dan; Dai, Wenjun; Gong, Jiaojiao; Li, Guande; Liu, Nansong; Wu, Wei; Pan, Jiaqiang; Chen, Chen; Jiao, Yingzhen; Deng, Huina; Ye, Junwei; Zhang, Xuanxuan; Huang, Huiling; Li, Qianyun; Xue, Liang; Zhang, Xiuming; Tang, Guanghui;  PLOS BIOL. 33320883 10.1371/journal.pbio.3000978

Rapid detection of SARS-CoV-2 with CRISPR-Cas12a.

Author(s):

Xiong, Dan; Dai, Wenjun; Gong, Jiaojiao; Li, Guande; Liu, Nansong; Wu, Wei; Pan, Jiaqiang; Chen, Chen; Jiao, Yingzhen; Deng, Huina; Ye, Junwei; Zhang, Xuanxuan; Huang, Huiling; Li, Qianyun; Xue, Liang; Zhang, Xiuming; Tang, Guanghui; 

Journal:

PLOS BIOL.

Year:

2020

Abstract:

The recent outbreak of betacoronavirus Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2), which is responsible for the Coronavirus Disease 2019 (COVID-19) global pandemic, has created great challenges in viral diagnosis. The existing methods for nucleic acid detection are of high sensitivity and specificity, but the need for complex sample manipulation and expensive machinery slow down the disease detection. Thus, there is an urgent demand to develop a rapid, inexpensive, and sensitive diagnostic test to aid point-of-care viral detection for disease monitoring. In this study, we developed a clustered regularly interspaced short palindromic repeats (CRISPR)-CRISPR associated proteins (Cas) 12a-based diagnostic method that allows the results to be visualized by the naked eye. We also introduced a rapid sample processing method, and when combined with recombinase polymerase amplification (RPA), the sample to result can be achieved in 50 minutes with high sensitivity (1-10 copies per reaction). This accurate and portable detection method holds a great potential for COVID-19 control, especially in areas where specialized equipment is not available.