RPB0294

Pathogen Description

Target Pathogen Pathogen Name NCBI Taxonomy ID Order Family Genus Species Pathogen type
Influenza A virus Influenza A virus, FLUAV, Human Influenza A Virus, Influenza virus type A 11320 Articulavirales Orthomyxoviridae Alphainfluenzavirus Influenza A virus 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
HA-RPA-F TCTAGTATGCCATTCCACAACATACACCCCC 31 10 µM 48.39 63.17 9304.12 \
HA-RPA-R AACCATCTACCATTCCCTGCCATCCTCCCTC 31 10 μM 54.84 66.83 9207.03 \

Gene Description

Target Gene GenBank ID
HA gene AF144305.1

Assay Description

Application Assay Primer Designing Software Reaction Time(min) Assay Temperature(℃) Readout System(s) Limit of Detection(LoD) Sensitivity(%) Specificity(%)
an accurate and convenient diagnostic tool, it can be used to diagnose the H5N1 avian influenza virus. RT-RPA DNAMAN 6.0 software 20 min 40 °C \ 1 × 10 -7 ng/μL 100 % 100 %

Publication Description

Year of Publication Title Author(s) Journal PMID DOI
2024 Dual Gene Detection of H5N1 Avian Influenza Virus Based on Dual RT-RPA Qian Wang,Shiwen Wu,Jiangbing Shuai,Ye Li,Xianshu Fu,Mingzhou Zhang,Xiaoping Yu,Zihong Ye,Biao Ma Molecules 38930866 10.3390/molecules29122801

Dual Gene Detection of H5N1 Avian Influenza Virus Based on Dual RT-RPA

Author(s):

Qian Wang,Shiwen Wu,Jiangbing Shuai,Ye Li,Xianshu Fu,Mingzhou Zhang,Xiaoping Yu,Zihong Ye,Biao Ma

Journal:

Molecules

Year:

2024

Abstract:

The H5N1 avian influenza virus seriously affects the health of poultry and humans. Once infected, the mortality rate is very high. Therefore, accurate and timely detection of the H5N1 avian influenza virus is beneficial for controlling its spread. This article establishes a dual gene detection method based on dual RPA for simultaneously detecting the HA and M2 genes of H5N1 avian influenza virus, for the detection of H5N1 avian influenza virus. Design specific primers for the conserved regions of the HA and M2 genes. The sensitivity of the dual RT-RPA detection method for HA and M2 genes is 1 × 10-7 ng/μL. The optimal primer ratio is 1:1, the optimal reaction temperature is 40 °C, and the optimal reaction time is 20 min. Dual RT-RPA was used to detect 72 samples, and compared with RT-qPCR detection, the Kappa value was 1 (p value < 0.05), and the clinical sample detection sensitivity and specificity were both 100%. The dual RT-RPA method is used for the first time to simultaneously detect two genes of the H5N1 avian influenza virus. As an accurate and convenient diagnostic tool, it can be used to diagnose the H5N1 avian influenza virus.