RPB0085

Pathogen Description

Target Pathogen Pathogen Name NCBI Taxonomy ID Order Family Genus Species Pathogen type
Coxsackievirus A6 CV-A6,CVA-6,,CVA6,Coxsackie A virus type 6,Enterovirus CA6,Human coxsackievirus A6,Human enterovirus CVA6 86107 Picornavirales Picornaviridae Enterovirus Enterovirus A Viruses

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
RT-RPA-F [Biotin]GGAGTTGTAGAGGTRAAGGACTCGGGYACTA 31 0.42 51.6 63.93 9704.85 \
RT-RPA-R TCCGGCTTAGGGGCYCCTGGTGGYACATACAT 32 0.42 59.4 71.33 9832.41 \
RT-RPA-P [FAM]AGGTTGGACACAAAAGTGAACTCRGCATCAA[dSpacer]GCGCATGTATGTTGA[C3-spacer] 46 0.12 44.6 68.68 14263.33 \

Gene Description

Target Gene GenBank ID
VP1 gene \

Assay Description

Application Assay Primer Designing Software Reaction Time(min) Assay Temperature(℃) Readout System(s) Limit of Detection(LoD) Sensitivity(%) Specificity(%)
Detection of Coxsackievirus A6 RT-RPA-LFS \ 35 37 LFS 10 copies per reaction \ 1

Publication Description

Year of Publication Title Author(s) Journal PMID DOI
2021 One-Step Reverse-Transcription Recombinase Polymerase Amplification Using Lateral Flow Strips for the Detection of Coxsackievirus A6 Jia Xie,Xiaohan Yang,Lei Duan,Keyi Chen,Pan Liu,Wenli Zhan,Changbin Zhang,Hongyu Zhao,Mengru Wei,Yuan Tang,Mingyong Luo Frontiers in microbiology 33613499 10.3389/fmicb.2021.629533

One-Step Reverse-Transcription Recombinase Polymerase Amplification Using Lateral Flow Strips for the Detection of Coxsackievirus A6

Author(s):

Jia Xie,Xiaohan Yang,Lei Duan,Keyi Chen,Pan Liu,Wenli Zhan,Changbin Zhang,Hongyu Zhao,Mengru Wei,Yuan Tang,Mingyong Luo

Journal:

Frontiers in microbiology

Year:

2021

Abstract:

Hand, foot, and mouth disease (HFMD) is a common infectious disease affecting mainly children under 5 years of age. Coxsackievirus A6 (CVA-6), a major causative pathogen of HFMD, has caused outbreaks in recent years. Currently, no effective vaccine or antiviral treatments are available. In this study, one-step reverse-transcription recombinase polymerase amplification (RT-RPA), combined with a disposable lateral flow strip (LFS) assay, was developed to detect CVA-6. This assay can be performed in less than 35 min at 37°C without expensive instruments, and the result can be observed directly with the naked eye. The sensitivity of the RT-RPA-LFS was 10 copies per reaction, which was comparable to that of the conventional real-time quantitative polymerase chain reaction (qPCR) assays. Moreover, the assay specificity was 100%. The clinical performance of the RT-RPA-LFS assay was evaluated using 142 clinical samples, and the coincidence rate between RT-RPA-LFS and qPCR was 100%. Therefore, our RT-RPA-LFS assay provides a simple and rapid approach for point-of-care CVA-6 diagnosis.