RPB0364

Pathogen Description

Target Pathogen Pathogen Name NCBI Taxonomy ID Order Family Genus Species Pathogen type
HRV Human rhinovirus 169066 Picornavirales Picornaviridae Enterovirus Human rhinovirus 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
HRV-B-F ATGTGCTTGRTTGWGANTCCTCCGGCCCCTGAATG 35 10 µM 54.29 69.46 10737.25 \
HRV-B-R GTWGANACYTGHGCDCCCATGRTCACAGTATAT 33 10 µM 45.94 62.88 10112.72 \

Gene Description

Target Gene GenBank ID
VP4 gene \

Assay Description

Application Assay Primer Designing Software Reaction Time(min) Assay Temperature(℃) Readout System(s) Limit of Detection(LoD) Sensitivity(%) Specificity(%)
a novel, rapid and simple method for detecting HRV-B infection based on isothermal nucleic acid-based amplification technology. RT-RPA-Cas12a BLAST 20 min 39 ℃ Cas12a 0.5 copies\µL \ \

Publication Description

Year of Publication Title Author(s) Journal PMID DOI
2023 Establishment of RT-RPA-Cas12a assay for rapid and sensitive detection of human rhinovirus B Yongdong Li,Xuefei Wang,Rong Xu,Ting Wang,Dandan Zhang,Weidong Qian BMC Microbiology 37951882 10.1186/s12866-023-03096-1

Establishment of RT-RPA-Cas12a assay for rapid and sensitive detection of human rhinovirus B

Author(s):

Yongdong Li,Xuefei Wang,Rong Xu,Ting Wang,Dandan Zhang,Weidong Qian

Journal:

BMC Microbiology

Year:

2023

Abstract:

Human rhinovirus B (HRV-B) is a major human viral pathogen that can be responsible for various kinds of infections. Due to the health risks associated with HRV-B, it is therefore crucial to explore a rapid, specific, and sensitive method for surveillance. Herein, we exploited a novel detection method for HRV-B by combining reverse-transcription recombinase polymerase amplification (RT-RPA) of nucleic acids isothermal amplification and the trans-cleavage activity of Cas12a. Our RT-RPA-Cas12a-based fluorescent assay can be completed within 35-45 min and obtain a lower detection threshold to 0.5 copies/µL of target RNA. Meanwhile, crRNA sequences without a specific protospacer adjacent motif can effectively activate the trans-cleavage activity of Cas12a. Moreover, our RT-RPA-Cas12a-based fluorescent method was examined using 30 clinical samples, and exhibited high accuracy with positive and negative predictive agreement of 90% and 100%, respectively. Taken together, a novel promising, rapid and effective RT-RPA-Cas12a-based detection method was explored and shows promising potential for on-site HRV-B infection in resource-limited settings.