RPB0353

Pathogen Description

Target Pathogen Pathogen Name NCBI Taxonomy ID Order Family Genus Species Pathogen type
Influenza A virus (H1N1) Influenza A virus (H1N1) 114727 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
H3-F CTGACTCAGAAATGAACAAACTGTTTGAAAA 31 10 μM 32.26 55.41 9544.32 \
H3-R GCAAAGGAAATCCATAGGATCCAATCTTTG 30 10 μM 40 57.62 9223.08 \
H3-P CTGAGGATATGGGCAATGGTTGTTTCAAAA(FAM-dT)(THF)(BHQ1-dT)ACCACAAATGTGACA [C3-spacer] 45 10 μM 40 65.42 13941.14 \

Gene Description

Target Gene GenBank ID
HA gene KT022317.1

Assay Description

Application Assay Primer Designing Software Reaction Time(min) Assay Temperature(℃) Readout System(s) Limit of Detection(LoD) Sensitivity(%) Specificity(%)
rapid, sensitive, specific, and cost-effective nature, combined with its potential for portability and POCT, make it an invaluable tool for early disease detection, control, and epidemiological surveillance RT-RAA SnapGene software (version 4.3.6) 30 min 42 °C \ 224 copies 0.9143 1

Publication Description

Year of Publication Title Author(s) Journal PMID DOI
2024 A Rapid Detection Method for H3 Avian Influenza Viruses Based on RT-RAA Jiaqi Li,Huan Cui,Yuxin Zhang,Xuejing Wang,Huage Liu,Yingli Mu,Hongwei Wang,Xiaolong Chen,Tongchao Dong,Cheng Zhang,Ligong Chen Animals 39272386 10.3390/ani14172601

A Rapid Detection Method for H3 Avian Influenza Viruses Based on RT-RAA

Author(s):

Jiaqi Li,Huan Cui,Yuxin Zhang,Xuejing Wang,Huage Liu,Yingli Mu,Hongwei Wang,Xiaolong Chen,Tongchao Dong,Cheng Zhang,Ligong Chen

Journal:

Animals

Year:

2024

Abstract:

The continued evolution of H3 subtype avian influenza virus (AIV)-which crosses the interspecific barrier to infect humans-and the potential risk of genetic recombination with other subtypes pose serious threats to the poultry industry and human health. Therefore, rapid and accurate detection of H3 virus is highly important for preventing its spread. In this study, a method based on real-time reverse transcription recombinase-aided isothermal amplification (RT-RAA) was successfully developed for the rapid detection of H3 AIV. Specific primers and probes were designed to target the hemagglutinin (HA) gene of H3 AIV, ensuring highly specific detection of H3 AIV without cross-reactivity with other important avian respiratory viruses. The results showed that the detection limit of the RT-RAA fluorescence reading method was 224 copies/response within the 95% confidence interval, while the detection limit of the RT-RAA visualization method was 1527 copies/response within the same confidence interval. In addition, 68 clinical samples were examined and the results were compared with those of real-time quantitative PCR (RT-qPCR). The results showed that the real-time fluorescence RT-RAA and RT-qPCR results were completely consistent, and the kappa value reached 1, indicating excellent correlation. For visual detection, the sensitivity was 91.43%, the specificity was 100%, and the kappa value was 0.91, which also indicated good correlation. In addition, the amplified products of RT-RAA can be visualized with a portable blue light instrument, which enables rapid detection of H3 AIV even in resource-constrained environments. The H3 AIV RT-RAA rapid detection method established in this study can meet the requirements of basic laboratories and provide a valuable reference for the early diagnosis of H3 AIV.