In this study, we assessed the efficiency of Pleurotus eryngii mushroom extract for killing larvae and attracting adult mosquito vectors (Aedes aegypti and Culex sitiens) in the Samut Songkhram province of Thailand. Five extract concentrations (120, 12, 1.2, 0.12 and 0.012 mg/L) were used for larvicidal tests, while 3 concentrations (100, 10, and 1 mg/L) were examined for adult mosquito attraction. The larvicidal results showed that P. eryngii extract did not kill Ae. aegypti larvae, while the extract had minimal effect on Cx. sitiens larvae except at 1.2 mg/L. P. eryngii extract most attracted adult Ae. aegypti and Cx. sitiens mosquitoes at 10 mg/L, followed by 1 and 100 mg/L, respectively. Further, statistical analysis revealed a significantly different number of mosquitoes that responded to P. eryngii extract and octenol. This research demonstrated that this mushroom extract could be developed to attract mosquitoes, although only one concentration (10 mg/L) attracted more than half of all Ae. aegypti adults.