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Discrimination between highly pathogenic avian influenza (HPAI) and low pathogenic avian influenza (LPAI) by melting curve analysis based on real-time reverse transcription–polymerase chain reaction of the H5 HA gene with SYBR Green I fluorescent dye. The melting peaks of HPAI and LPAI were clearly separated. The cutoff value was set at 78.50°C (midpoint between HPAI and LPAI) and used to interpret the pathogenicity of unknown samples.
