Based on known ITS sequences of the rice cyst Nematode (Heterodera elachista) and common cyst nematodes, one species-specific primer pair named HeF and HeR was designed for H. Elachista, the expected fragment length of the PCR products using the primers was 281 bp. Using the established DNA extraction method and PCR system with the specific primers, target fragment detected from a single second instar larvae of nematode or an individual H. elachista mixed in 0.1 g rice root tissues. Combined the species-specific primers He-F and He-R with universal primer pairs D2A and D3B, a duplex PCR was established, which detects H. elachista from cyst nematode samples primarily separated from field soil.