RPB0435

Pathogen Description

Target Pathogen Pathogen Name NCBI Taxonomy ID Order Family Genus Species Pathogen type
MERS-CoV Middle East respiratory syndrome-related coronavirus, MERS coronavirus, MERS virus, Middle East Respiratory Syndrome Coronavirus, Middle East respiratory syndrome coronavirus 1335626 Nidovirales Coronaviridae Betacoronavirus Nidovirales 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
MERS-upE-F CTCTGCGCTACTATGGGTCCCGTGTAGAG 29 10 μM 58.62 65.83 8900.81 \
MERS-upE-R CTCTACACGGGACCCATAGTAGCGCAGAG 29 10 μM 58.62 65.83 8896.84 \

Gene Description

Target Gene GenBank ID
upE \

Assay Description

Application Assay Primer Designing Software Reaction Time(min) Assay Temperature(℃) Readout System(s) Limit of Detection(LoD) Sensitivity(%) Specificity(%)
a strong supplement in the prevention and early diagnosis of infectious diseases in resource-limited situations. RT-RAA-CRISPR Cas12a Primer Premier 6 and primer-BLAST 30 min 42 °C CRISPR Cas12a 1 copy per μL 1 0.908

Publication Description

Year of Publication Title Author(s) Journal PMID DOI
2023 CRISPR Cas12a-enabled biosensors coupled with commercial pregnancy test strips for the visible point-of-care testing of SARS-CoV-2 Mycoplasma pneumonia Peijie Shen,Zhenjun Si,Di Huang,Zhipeng Xu,Ziyi Wang,Mengjun Fang,Zhinan Xu Analyst 37159023 10.1039/d3an00284e

CRISPR Cas12a-enabled biosensors coupled with commercial pregnancy test strips for the visible point-of-care testing of SARS-CoV-2 Mycoplasma pneumonia

Author(s):

Peijie Shen,Zhenjun Si,Di Huang,Zhipeng Xu,Ziyi Wang,Mengjun Fang,Zhinan Xu

Journal:

Analyst

Year:

2023

Abstract:

The rapid spread of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has promoted the development of nucleic acid diagnosis technology. Several platforms with isothermal amplification methods have achieved sensitive and specific detection of SARS-CoV-2. However, they still suffer from complicated operations, delicate instruments, and unintuitive signal output modes. Here, a system consisting of CRISPR Cas12a-based biosensors and commercial pregnancy test strips (CRISPR-PTS) was established for the point-of-care testing of SARS-CoV-2. The target viral nucleic acids were finally reflected on the test strips through four steps, namely sample pretreatment, RT-RAA amplification, CRISPR Cas12a reaction, and separation-free hCG detection. This CRISPR-PTS assay possessed an outstanding sensitivity of as low as 1 copy per μL for SARS-CoV-2 detection and showed an excellent specificity in distinguishing the SARS-CoV-2 pseudovirus as well as other SARS-like viral clinical samples. In addition, the CRISPR-PTS assay performed well in practical applications, with 96.3% agreement versus RT-qPCR in spiked samples. With the advantages of low reagent cost, simple operation procedure, and visible signal output, CRISPR-PTS assay was expected to provide a strong supplement in the prevention and early diagnosis of infectious diseases in resource-limited situations.