Spodoptera litura Fabricius is the serious, polyphagous pest of various crops. Due to the high resistant to most of chemical pesticides, it is hard to control S. litura in the fields. We investigated the virulence of four entomopathogenic nematodes (EPNs) in each larval stage. All EPNs were more effective to younger larvae than older larvae. Mortality, larval penetration rate and reproduction rate were significantly higher in H. indica, S. carpocapsae and S. longicaudum than the H. bacteriophora. Three high effective EPNs induced complete mortality of first instar larvae within 48 h, but of fifth instar larvae within 72 h. These EPNs also highly effective to control S. litura in pot assay. Thus, H. indica, S. carpocapsae and S. longicaudum, can be used as efficient biological control agents of S. litura.