Reverse transcription-polymerase chain reaction (RT-PCR)
The full-length of the vp60gene sequence was amplified by RT-PCR using the Reverse Transcriptase XL
(AMV) kit (Takara Bio, Dalian, China) and the Ex Taq kit (Takara Bio). A
specific primer pair (sense, 5′-AAGAGAGTCGTCTCGGTAGTA-3′ and antisense
5′-GCGCCTGCAAGTCCCAATCC-3′) was used as described previously (Duarte et
al., 2014). The vp60 gene was then cloned into a pMD-19T vector
(Takara Bio). Positive clones were sequenced and analyzed further.