Vaccinia virus is a ubiquitous DNA virus in nature. It can express exogenous genes efficiently and induce both humoral and cellular immunity. This property has made it become a hot topic in gene therapy research. Since the probability of homologous recombination is low, effective screening and purification of vaccinia virus is very important. At present, the commonly used methods for screening vaccinia virus are based on the use of reporter genes, such as GFP, LacZ, gpt, and other genes, or the use of TK^- and Brdu combined with fluorescence. Some scholars have tried to fuse drug resistance gene, fluorescence, and chromogenic genes to carry out drug screening and observe gene expression. In this article, we will make a summary of the commonly used methods for vaccinia virus screening and purification, and compare their advantages and disadvantages.