Abstract: To quickly identify entomopathogenic nematodes(EPN) and confirm the difference among species/strains of EPNs kept in our laboratory for many years,the conserved ribosome DNA(rDNA,18S-ITS1-5.8S-ITS2-28S) region of EPN was used for PCR amplification and sequencing.The comparison and analysis of 18S-28S rDNA region and 28S rDNA D2-D3 extension segments of two genera of Heterorhabditis and Steinernema including 13 EPN species/strains were conducted to identify the similarity of these nematodes with reported nematodes in GenBank.The results of molecular identification with 28S rDNA D2-D3 region were fully matched with those from morphological data and the similarity with reported nematodes in the GenBank reached up to 99%-100%.The D2-D3 region sequences clearly distinguished EPN genus,species or strains.The amplification region of 18S-ITS1-5.8S rDNA separated S.glaseri and S.carpocapsae but not for closely related S.litorale and S.feltiae.The internal transcribed spacer ITS2 rDNA would differentiate S.litorale and S.feltiae.The study indicated rDNA would be utilized for rapid identification of EPN species/strains.