[Background] The biological activity of Bacillus thuringiensis (Bt) strains collected from soil samples was tested against Opisina arenosella Walker, in order to obtain high insect activity strains that may help control the species. The second objective of this study was to provide new genetic resources for transgenic plant of Bt that could be more resistant to this species. [Method] Soil samples were collected from different areas of Hainan Province potentially suitable for O. arenosella. The temperature screening method was used to isolate Bt strains, which were then identified based on their morphology and molecular biology. The toxicity of Bt strains to larvae of O. arenosella was tested by leaf immersion bioassay method. [Result] Eight Bt isolates were obtained from Baoting soil samples, Hainan Province. The 8 Bt isolates included bipyramidal crystal, spherical crystal, square crystal types. PCR-RFLP analysis showed that Cry1, cry1I and cry2 type genes were present in these isolates, but the cry gene-types of two isolates were unknown. The toxicity of the extracted insecticidal crystal proteins (ICP) of the 8 strains against larvae of O. arenosella was assayed. The results indicated that the use of ICP of BAT10 (50 μg·mL^-1) resulted in 100% mortality of the larvae. ICP of 29-15-4 and BAT20 resulted in more than 80% mortality while the ICP of the other five strains showed less than 50% mortality. [Conclusion and significance] This study showed that effective Bt strains could be found in soil potentially suitable to O. arenosella and among them, two strains had the potential to be used as biological control against this species.