African swine fever (ASF) is a highly contagious and deadly viral disease in domestic and feral pigs. Since 2018, the disease has spread and caused large socioeconomic consequences to the pig industry in several Asian countries including China, Vietnam, and South Korea. This study aims to determine the genotype, serotype, and genetic variation of representative ASF viruses (ASFV) responsible for the outbreaks in 2019–2022 in Can Tho city, a central administrative province in the Mekong delta, Vietnam. For outbreak investigation, the presence of causative ASFVs was tested using conventional PCR targeting the B646L gene. Subsequently, the amplification and sequencing of the DNA fragments of the putative B646L gene encoding the major capsid protein p72, EP402R gene encoding the viral hemagglutinin CD2-like protein (CD2v), and intergenic region (IGR) between the l73R and I329L genes were performed for molecular characterization. Phylogenetic analyses based on B646L and EP402R genes confirmed that all ASFVs detected in Can Tho city belonged to genotype 2 and serotype 8. In addition, this study revealed that at least two variants of ASFVs, namely, IGR II and IGR III, based on the nucleotide variation of the IGR sequence, cocirculated, and caused outbreaks in Can Tho city. The molecular characterization study provides great significance for understanding the evolution of ASFVs and tracing possible sources of infection in Can Tho and Mekong delta.