Klebsiella spp. are an important environmental pathogen of mastitis in dairy cows, which cause many severe clinical symptoms and great economical losses of dairy farms. The aim of the work was to identify the prevalence of Klebsiella spp. from bovine mastitis milk in China, antibiotic resistance phenotypes, and the main virulence genes. We investigated 497 milk samples collected from cow with clinical mastitis in dairy farms (located in 5 provinces) of China in 2017. The minimum inhibitory concentrations (MICs) of antimicrobial susceptibility testing were determined for Klebsiella spp. isolates, the assay was performed by BD Phoenix 100 and the results were interpreted by the guidance of Clinical and Laboratory Standards Institute (CLSI, 2016) as susceptible, intermediate, and resistant. The detection of resistance genes and virulence genes was performed by real-time polymerase chain reaction. In our study, 32 (6.4%) isolated from 497 samples were identified as Klebsiella spp. An analysis of antimicrobial susceptibility testing showed that the highest percentage of the isolates were resistant to Tetracycline (40.6%). None isolates were resistant to Piperacillin-Tazobactam, Levofloxacin, Fosfomycin w/G6P. An analysis of the prevalence of antimicrobial resistance genes revealed that Sul2 gene was the most prevalent among all the resistance genes (96.9%), followed by blaOXA-48 (75%). Further investigation of virulence genes performed by PCR found that all the strains harbored one or more of these virulence genes, the percentage of the isolates were positive for ureA was 100%. The present study demonstrated that the prevalence of resistance and virulence genes was high, and it could contribute a threat to the health of animals or even humans. The result of the study highlighted the need for improvements in antimicrobial therapy and infection control programs in Chinese dairy herds to reduce the prevalence of resistance of Klebsiella spp.