A sample of Aedes aegypti (L.) from Santiago de Cuba, Cuba, with a high level of temephos resistance (19.58× at the 50% lethal concentration [LC₅₀]), was subjected to temephos selection to evaluate the utility of this organophosphate insecticide for mosquito control. High resistance developed after six generations of selection (200.00×). Little or no cross-resistance was observed to the organophosphates, malathion and fenitrothion, but high cross-resistance was observed for the pyrethroid deltamethrin (337.5×) and the organophosphate fenthion (12.74×). Synergism tests implicated detoxifying esterases in temephos and fenthion resistance and deltamethrin resistance was associated with a cytochrome P450 monooxygenasa. Biochemical tests, polyacrylamide gel electrophoresis (PAGE), and inhibition studies confirmed the presence of elevated esterase activity associated with temephos resistance. Pyrethroid resistance could be associated with a glutathione-S-transferase mechanism but not the esterases. The cross-resistance to deltamethrin from temephos selection could limit the use of both insecticides for Ae. aegypti control.