[Objectives]  Dysmicoccus neobrevipes Beardsley, a new invasive species in China, is a worldwide pest that poses a serious threat to agriculture and forestry. Morphological identification of this species is limited by its close similarity to other mealybug species. [Methods]  To address this problem a new method for the rapid identification of D. neobrevipes was developed and a pair of 28S rDNA species-specific primers (D.neo-28SF/R) were designed. The 28S rDNA genes of D. neobrevipes and six other mealybug species were amplified and sequenced. [Results]  28S rDNA species-specific primers (D.neo-28SF/R) amplified a single band of 235 bp for D. neobrevipes. The species-specificity of this primer pair was validated with the six other mealybug species mentioned above. The results show that only D. neobrevipes rDNA was amplified, and no products from other mealybugs were obtained. The method was also tested on individual D. neobrevipes from different countries and hosts. [Conclusion]  The diagnostic PCR assay established here provides a quick, simple and reliable molecular technique for the identification of D. neobrevipes, which will be useful in intercepting and preventing further spreading of this pest.