RPB0421

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
RT-RAA F3 GAACCTTTATGACAAAGTCCGACTACAGC 29 10 µM 44.83 59.08 8854.84 \
RT-RAA R1 TTCTGAGTACTGGGGGTAGTCATACGTTC 29 10 µM 48.28 60.88 8954.86 \
H5-LFD Probe FAM-TAGGGATAATGCAAAGGAGCTGGGTAATGGTT[THF]TTTCGAGTTCTATC -3′C3spacer 46 10 µM 41.3 65.14 14314.34 \

Gene Description

Target Gene GenBank ID
HA gene \

Assay Description

Application Assay Primer Designing Software Reaction Time(min) Assay Temperature(℃) Readout System(s) Limit of Detection(LoD) Sensitivity(%) Specificity(%)
a swift, portable, and precise method of determining results for the detection of H5-AIV. RT-RAA-LFD \ 20 min 39°C LFD 1 copy/μL 0.85 100 %

Publication Description

Year of Publication Title Author(s) Journal PMID DOI
2023 Establishment of two assays based on reverse transcription recombinase-aided amplification technology for rapid detection of H5 subtype avian influenza virus Yang Li,Jiajing Shang,Yixin Wang,Juan Luo,Wenming Jiang,Xin Yin,Fuyou Zhang,Chunran Deng,Xiaohui Yu,HuaLei Liu Microbiology Spectrum 37811963 10.1128/spectrum.02186-23

Establishment of two assays based on reverse transcription recombinase-aided amplification technology for rapid detection of H5 subtype avian influenza virus

Author(s):

Yang Li,Jiajing Shang,Yixin Wang,Juan Luo,Wenming Jiang,Xin Yin,Fuyou Zhang,Chunran Deng,Xiaohui Yu,HuaLei Liu

Journal:

Microbiology Spectrum

Year:

2023

Abstract:

Avian influenza virus (AIV) subtype H5 is a highly contagious zoonotic disease and a serious threat to the farming industry and public health. Traditional detection methods, including virus isolation and real-time PCR, require tertiary biological laboratories and are time-consuming and complex to perform, making it difficult to rapidly diagnose H5 subtype avian influenza viruses. In this study, we successfully developed two methods, namely, RF-RT-RAA and RT-RAA-LFD, for rapid detection of H5-AIV. The assays are characterized by their high specificity, sensitivity, and user-friendliness. Moreover, the results of the reaction can be visually assessed, which are suitable for both laboratory testing and grassroots farm screening for H5-AIV.