Abstract
BackgroundRecent epidemiological investigation of different HA subtypes of avian influenza viruses (AIVs) shows that the H3 subtype is the most predominant among low pathogenic AIVs (LPAIVs), and the seasonal variations in isolation of H3 subtype AIVs are consistent with that of human H3 subtype influenza viruses. Consequently, the development of a rapid, simple, sensitive detection method for H3 subtype AIVs is required. The loop-mediated isothermal amplification (LAMP) assay is a simple, rapid, sensitive and cost-effective nucleic acid amplification method that does not require any specialized equipment.ResultsA reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay was developed to detect the H3 subtype AIVs visually. Specific primer sets target the sequences of the hemagglutinin (HA) gene of H3 subtype AIVs were designed, and assay reaction conditions were optimized. The established assay was performed in a water bath for 50 minutes, and the amplification result was visualized directly as well as under ultraviolet (UV) light reflections. The detection limit of the RT-LAMP assay was 0.1pg total RNA of virus, which was one hundred-fold higher than that of RT-PCR. The results on specificity indicated that the assay had no cross-reactions with other subtype AIVs or avian respiratory pathogens. Furthermore, a total of 176 clinical samples collected from birds at the various live-bird markets (LBMs) were subjected to the H3-subtype-specific RT-LAMP (H3-RT-LAMP). Thirty-eight H3 subtype AIVs were identified from the 176 clinical samples that were consistent with that of virus isolation.ConclusionsThe newly developed H3-RT-LAMP assay is simple, sensitive, rapid and can identify H3 subtype AIVs visually. Consequently, it will be a very useful screening assay for the surveillance of H3 subtype AIVs in underequipped laboratories as well as in field conditions.
Highlights
Recent epidemiological investigation of different HA subtypes of avian influenza viruses (AIVs) shows that the H3 subtype is the most predominant among low pathogenic AIVs (LPAIVs), and the seasonal variations in isolation of H3 subtype AIVs are consistent with that of human H3 subtype influenza viruses
There is no evidence that H5N1 HPAIVs can transmit widely between humans; the human-pandemic influenza strains are all generated by gene reassortment between human influenza viruses, avian influenza viruses and swine influenza viruses [7]
It’s notable that the results of a recent epidemiological investigation into different HA subtype AIVs shows that the H3 subtype is the predominant subtype among LPAIVs, and the seasonal variations in isolation of H3 subtype AIVs are consistent with that of human H3 subtype influenza viruses [8]
Summary
Recent epidemiological investigation of different HA subtypes of avian influenza viruses (AIVs) shows that the H3 subtype is the most predominant among low pathogenic AIVs (LPAIVs), and the seasonal variations in isolation of H3 subtype AIVs are consistent with that of human H3 subtype influenza viruses. The development of a rapid, simple, sensitive detection method for H3 subtype AIVs is required. It’s notable that the results of a recent epidemiological investigation into different HA subtype AIVs shows that the H3 subtype is the predominant subtype among LPAIVs, and the seasonal variations in isolation of H3 subtype AIVs are consistent with that of human H3 subtype influenza viruses [8]. It is important to enhance surveillance for H3 subtype AIVs infections and to develop a simple, rapid and sensitive detection method
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