Simple SummaryPork is one of the most important types of sources of protein in the human diet. Swine diseases cause major economic losses to the global pig industry. Early diagnosis and prevention of diseases are fundamental factors for maintaining the health of animals on pig farms. The present article is the first report on the successful isolation of the swinepox virus in Russia. The results of a laboratory experiment on animals revealed a low pathogenicity of the Russian isolate for pigs. We suggest that this isolate can be used as a viral vector for the development of vaccines for animals. In addition, we have demonstrated for the first time that nasal and oral swabs can be used for PCR diagnosis of the disease and for swinepox virus isolation.AbstractSwinepox virus (SWPV) is the only member of theSuipoxvirusgenus of thePoxviridaefamily and is an etiologic agent of a worldwide disease specific for domestic and wild pigs. SWPV outbreaks are sporadically recorded in different regions of Russia. In 2013, an outbreak of the disease causing skin lesions was registered on a pig farm in Russia. The presence of SWPV in the scab samples was assessed by in-house real-time PCR, reference PCR amplification, and nucleotide sequencing of the viral late transcription factor-3 (VLTF-3) gene and was then confirmed by virus isolation. Thus, the in-house real-time PCR proposed in this study could serve as a useful tool for the rapid specific detection of the swinepox virus. In the study, it has been demonstrated for the first time that nasal and oral swabs can be used for PCR diagnosis of the disease and for swinepox virus isolation. Phylogenetic analysis revealed that the isolated virus was closely related to SWPV isolates registered in Germany, USA, and Brazil, and slightly differed from the Indian isolates. During experimental infection of pigs, a low pathogenicity of the Russian isolate was observed. Our data provides the first report on the isolation and characterization of swinepox virus in Russia.