[Aim] This study aimed to examine the infection effect of entomopathogenic nematode on the 2^nd and 5^th instar larvae of Spodoptera frugiperda (J.E. Smith).[Method] We used a bioassay to test the virulence of four entomopathogenic nematodes:Steinernema carpocapsae AⅡ, S. longicaudum X-7, S. feltiae SN and Heterorhabditis bacteriophora H06, under laboratory conditions.[Result] After exposing one S. frugiperda larva to 50 S. carpocapsae AⅡ for 36 hours, the mortality rates were 92% for 2^nd and 100% for 5^th instar S. frugiperda larvae. When exposed to 50 S. longicaudum X-7, the mortality of 2^nd instar S. frugiperda larvae was 80% and 100% for 5^th instar S. frugiperda larvae in the presence of only 30 S. longicaudum X-7. After exposing one S. frugiperda larva face to 400 S. feltiae SN for 36 hours, the mortality rates of 2^nd and 5^th instar S. frugiperda larvae were 88% and 85%and 32% and 67.5% in the presence of 400 H. bacteriophora H06.[Conclusion] The results showed that S. carpocapsae AⅡ and S. longicaudum X-7 had great potential on biological control of 2^nd and 5^th instar S. frugiperda larvae.