BACKGROUND: While initial Human Papillomavirus (HPV) vaccine uptake estimates are encouraging, especially in the school-aged population, it is unclear to what extent general practitioners (GPs) have been recommending and administering the vaccine to adult women and what the uptake has been. METHODS: A mixed methods study of Australian GPs consisting of a knowledge and attitudes questionnaire and an audit which assessed overall vaccination rates since commencement of the Australian National HPV program and in the last 50 female patients aged 27-45 years who consulted them. RESULTS: GPs have a good level of knowledge regarding HPV vaccination and are strongly committed to it for women until 27 years but are much less so for women aged 27-45 years. From 2007 to 2010, only 1.9% of women aged 27-45 who consulted the GPs commenced HPV vaccination. Of those, however, the majority completed all three doses. Higher rates of commencement of HPV vaccination (11.4%) were demonstrated by auditing the last 50 consecutive patients seen by the GP. In this group of women, 63% had received a Pap test in the last two years. Female GPs had significantly higher rates of vaccination. CONCLUSIONS: There is relatively low HPV vaccine uptake in women aged 27-45. Once HPV vaccination is commenced, however, completion rates are high in women in this age group. Low uptake may be due to lack of opportunistic awareness raising in relevant consultations. Clear guidance together with further exploration of patient factors and GP barriers and enablers would assist implementation of HPV vaccination.