Viral RNA Extraction and RT-PCR
The HA-positive allantoic fluid was used for viral RNA extraction using
a High Pure Viral Nucleic Acid Kit according to the manufacturer’s
instructions (Roche, Mannheim, Germany). Reverse transcription was
performed with the Uni12 primer (5ʹ-AGCAAAAGCAGG-3ʹ) and the RevertAid
First-Strand cDNA Synthesis Kit (Fermentas, Canada). cDNAs of the eight
segments were then amplified with gene-specific primers designed by the
CNIC. All experiments were conducted in accordance with a previous
published study[15].