The Australian National Poliovirus Reference Laboratory was established in late 1994, as part of Australia's commitment to the World Health Organization's (WHO) polio eradication program. The laboratory continues to play a pivotal role in maintaining Australia's polio-free status through surveillance for cases of acute flaccid paralysis (AFP), the main clinical presentation of poliomyelitis, and the testing of specimens from these cases. The annual notification rate for eligible cases of AFP in Australia for 2003 was 0.83 per 100,000 children less than 15 years of age. The annual non-polio AFP rate after classification of cases by the polio expert committee was 0.68 per 100,000, 32 per cent below WHO's annual target. While no polioviruses were isolated from the specimens tested from the 27 cases of AFP in 2003, a novel enterovirus (enterovirus 75) was isolated from one case and enterovirus 71 was isolated from another. During the same period 12 polioviruses, referred from cases other than AFP, tested as Sabin-like by the WHO approved methods of intratypic differentiation. The importation of wild polioviruses from endemic Nigeria into surrounding countries of Africa during 2003, highlights the importance of the continuation of AFP surveillance and high quality laboratory activities throughout the world until global eradication of polio is certified.