Establishment of a reverse transcription loop-mediated isothermal amplification assay for detection of H6 subtype avian influenza virus

Abstract:

To develop a rapid sensitive and specific method of a reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay for detection of H6 subtype avian influenza virus (AIV), six primers specific to the eight site of HA gene were designed according to the sequences of H6 subtype AIV hemagglutinin gene available in GenBank. The viral RNA was amplified with one-step RT-LAMP method, and the reaction conditions were optimized. The results showed that the specificity of the assay was fine without cross amplification of other respiratory tract pathogens, and the detection limit of the RT-LAMP assay was 0.01 pg. The assay is a sensitive, simple, rapid and practical method for detection of H6 subtype AIV in the field.