RPB0388

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
AIV-F CTCATG GARTGG MTAA AGACAAGAC CAAT 29 10 μM 41.38 58.25 8947.91 \
AIV-R ACAAAVCGTCTACGC TGCAGTCCTCGC TCA 30 10 μM 55.57 68.22 9093.3 \
AIV-P CAC CTC TGACTA ARG GGATTT TAG GRT TTG \i6FAMdT\ \THF\ \iBHQ1dT\TCACGC TCACC[C3-spacer 41 10 μM 48.78 67.16 12551.19 \

Gene Description

Target Gene GenBank ID
M gene KU042441.1

Assay Description

Application Assay Primer Designing Software Reaction Time(min) Assay Temperature(℃) Readout System(s) Limit of Detection(LoD) Sensitivity(%) Specificity(%)
an effective and practicable tool for AIV molecular detection RT-RAA Oligo 7 20 min 39°C \ 102RNA copies 0.9759 0.9679

Publication Description

Year of Publication Title Author(s) Journal PMID DOI
2023 Reverse transcription recombinase-aided amplification assay for avian influenza virus Suchun Wang,Qingye Zhuang,Nan Jiang,Fuyou Zhang,Qiong Chen,Ran Zhao,Yang Li,Xiaohui Yu,Jinping Li,Guangyu Hou,Liping Yuan,Fuliang Sun,Zihao Pan,Kaicheng Wang Virus Genes 36781819 10.1007/s11262-023-01979-z

Reverse transcription recombinase-aided amplification assay for avian influenza virus

Author(s):

Suchun Wang,Qingye Zhuang,Nan Jiang,Fuyou Zhang,Qiong Chen,Ran Zhao,Yang Li,Xiaohui Yu,Jinping Li,Guangyu Hou,Liping Yuan,Fuliang Sun,Zihao Pan,Kaicheng Wang

Journal:

Virus Genes

Year:

2023

Abstract:

Avian influenza virus (AIV) infection can lead to severe economic losses in the poultry industry and causes a serious risk for humans. A rapid and simple test for suspected viral infection cases is crucial. In this study, a reverse transcription recombinase-aided amplification assay (RT-RAA) for the rapid detection of all AIV subtypes was developed. The reaction temperature of the assays is at 39 °C and the detection process can be completed in less than 20 min. The specificity results of the assay showed that this method had no cross-reaction with other main respiratory viruses that affect birds, including Newcastle disease virus (NDV) and infectious bronchitis virus (IBV). The analytical sensitivity at a 95% confidence interval was 102 RNA copies per reaction. In comparison with a published assay for reverse transcription quantitative real-time polymerase chain reaction (RT-qPCR), the κ value of the RT-RAA assay in 384 avian clinical samples was 0.942 (p < 0.001). The sensitivity and specificity of the RT-RAA assay for avian clinical sample detection was determined as 97.59% (95% CI 93.55-99.23%) and 96.79% (95% CI 93.22-98.59%), respectively. The RT-RAA assay for AIV in this study provided an effective and practicable tool for AIV molecular detection.