DNA barcoding is a recently developed technique for species-level identification that involves the use of short, standard DNA sequences as species labels. It is an effective complement to traditional taxonomic classification based on morphology. At present, research and applications involving the DNA barcoding of the Pseudococcidae are focused primarily on the cytochrome coxidase subunit I (COI) gene, but there is not yet a consensus on the preferred gene region for barcoding. The purpose of this study was to explore the effectiveness of identification of Pseudococcidae beetles using DNA barcoding technology. The COI gene sequences of 97 samples from 21 species of Asemini were analysed, followed by evaluation of the ability to identify species using a tree-building method and distance evaluation. The COI sequences (500 bp) exhibited distinct distributions of intra-specific and inter-specific variation and a significant barcoding gap. The success rate of identification was 97.84%. These results demonstrate the feasibility of using this segment of COI to identify most species of Pseudococcidae.