Spodoptera litura (F.) is an obnoxious cosmopolitan pest that causes serious damage to different economic crops. Entomopathogenic nematodes (EPN) have the potential to control the S. litura larvae. Fifteen EPN isolates were screened, and Steinernema sp. 64-2, four isolates of S. carpocapsae (Weiser), S. longicaudum (Shen & Wang) X-7, and two isolates of H. indica (Poinar, Karunaka & David) were found to cause higher mortality of the second, third, and fourth instars of S. litura than the other tested isolates, with larval mortality rates > 90% after 48 h of exposure. An exposure rate of 12.5 infective juveniles per larva was enough for S. carpocapsae A24, All, and G-R3a-2 and S. longicaudum X-7 to cause 100% mortality of the second instar, and for S. longicaudum X-7 and H. indica 212-2 to cause 100% mortality of the third instar. Five EPN isolates were tested on their virulence at different temperatures and found that all the five EPN isolates performed well against the S. litura larvae at 25 and 30°C, but were not active at 10 and 15°C. Two S. carpocapsae isolates (All and Mex) were virulent against the S. litura larvae at lower temperatures. The five tested EPN isolates were also found to have the ability to infect and kill the pupae of S. litura in the laboratory. The present study further proves that EPN are effective at controlling S. litura, which may partially substitute the use of chemical insecticides, thus reduce the overuse of chemical insecticides.