The development of insecticide resistance has compromised mosquito control efforts in many parts of the world. Papua New Guinea (PNG) has a long history of dichlorodiphenyltrichloroethane (DDT) use and currently distributes pyrethroid-treated nets for malaria control. This study is the first to investigate the status of pyrethroid resistance in the Anopheles punctulatus group, the major malaria and filariasis vectors of PNG. The study used World Health Organization standard susceptibility bioassays to detect knockdown phenotypes and a novel nested polymerase chain reaction to detect the knockdown resistant (kdr) allele in these vectors. Our results show 100% susceptibility to pyrethroids in all populations surveyed and an absence of the kdr allele.