Glycosyltransferases (GTs) are a diverse group of transferases widely distributed in microorganisms and plants which catalyze the transfer of glycosyl moieties from activated sugar donors to a diverse range of acceptor molecules. GTs play important roles in biological processes, such as cell signal transduction, cell-cell interactions, immune response and diseases. In addition, the utilization of glycosyltransferases to synthesize glycosides has become a major area of pharmaceutical research. The construction of a rapid and efficient screening method for glycosyltransferases is the key to the successful discovery, identification, and modification of new enzymes. In the past few decades, a series of screening methods have been developed to assess activity. This review focuses on various methods for rapid screening of GTs, outlining their advantages and disadvantages.