Emergence and molecular characterization of bovine papular stomatitis virus in Chinese cattle

The serum and swab samples were centrifuged at 4 ℃ at 12,000 ×g for 5 min. The supernatants were then filtered through 0.45 µm-pore-sized membranes (Merck Millipore, Taufkirchen, Germany), and digested with DNase I and RNase A (Takara, Shiga, Japan) to eliminate the contamination of foreign nucleic acids (NAs). Total RNA was extracted from pooled samples using the TRIzol reagent (Invitrogen), followed by ribosomal RNA (rRNA) depletion.

Subsequently, the NEBNext Super-Directed RNA Library Prep Reagents cassette (New England Biolabs, Ipswich, USA) was employed for meta-transcriptomic (MTT) library construction. For multiple displacement amplification (MDA) library preparation, DNA was extracted using the DNeasy Blood and Tissue Kit (Qiagen, Hilden, Germany) and amplified using the illustra GenomiPhi V2 DNA Amplification Kit (GE, Fairfield, USA). The amplified products were then purified using the QIAquick PCR purification Kit (Qiagen, Hilden, Germany), followed by Illumina paired-end sequencing (150 bp) on an Illumina NovaSeq 6000 sequencer.The viral sequences of the Bovine papular stomatitis virus，Influenza virus D, Rotavirus A and Bovine rhinitis A virus were assembled using MEGAHIT software.All contigs have been uploaded.