Insecticide resistance due to mutations in the knockdown resistance (kdr) gene in Anopheles gambiae populations has been reported in some West African countries, despite the absence of large-scale mosquito control in this sub-region. Agricultural and/or household use of insecticides has been implicated in the development of resistance in An. gambiae. However, such studies had not been carried out in Ghana, where large-scale use of insecticide-treated bed nets is a major component of the Roll Back Malaria programme. There is therefore a need to establish baseline information on patterns of insecticide use and vector susceptibility levels in Ghana. A questionnaire survey of 171 households with 800 inhabitants was conducted to obtain information on knowledge, attitude and perception of respondents on household insecticide use in the Accra metropolis. Also, adult An. gambiae were tested for resistance to permethrin and deltamethrin using WHO adult mosquito test kits. The polymerase chain reaction (PCR) was used to detect the presence of the kdr mutation in the test mosquitoes and to identify sibling species within the An. gambiae complex. Results indicated a high frequency of household pyrethroid insecticide use with respondents preferring aerosols to insecticide-impregnated coils. The use of anti-mosquito screens on doors and windows was very high (77 %) but use of bed nets was low (5 %). There was no significant difference in the reported incidence of malaria in all socioeconomic classes. All of the mosquito specimens examined were found to be An. gambiae s.s. Bioassay mortalities were 76 % (range: 72–80 %) for deltamethrin and 74.74 % (range 68–85 %) for permethrin, indicating the existence of resistance to these two insecticides in An. gambiae populations in Accra. The kdr mutation was found in 81.3 % of the survivors of the bioassays and in 8.8 % of the test mortalities. There was a strong correlation between resistance and the occurrence of kdr.