The invention discloses a complete genome sequence and an amplification primer of a Seneca valley virus SVV/CH/NM/2016. A one-step RT-PCR (Reverse Transcription-Polymerase Chain Reaction) method is firstly adopted for amplifying seven nucleotide sequence fragments (S1, S2, S3, S4, S5, S6, and S7) of an SVV/CH/NM/2016 strain, then the seven nucleotide sequence fragments are cloned and sequenced, and then, the DNA sequences of the seven nucleotide sequence fragments are sequentially spliced, edited and corrected to finally obtain the complete genome sequence of the Seneca valley virus SVV/CH/NM/2016. The obtained complete genome sequence of the Seneca valley virus SVV/CH/NM/2016 is favorable for further research of the pathogenic mechanism of the Seneca valley virus, molecular epidemiology,reverse genetics and the like, thereby establishing important data support and theoretical basis for diagnostic reagent development, vaccine development and the like of the Seneca valley virus.