Needlestick Injuries (NSI) commonly occurs in the hospital setting. Medical students are at risk of NSI due to lack of clinical experience. The purpose of this study was to investigate the incidence and knowledge, attitudes, awareness and practices of NSI among medical students at an Australian university. We conducted a cross-sectional study using an online questionnaire on NSI-related knowledge, attitude, awareness, practice, incidents and follow up. The survey was distributed to clinical students (years 3-6) of a 6-year medical program in Australia. Data were analysed with descriptive and bivariate analyses (IBM SPSS v.22; alpha 0.05). Response rate to the study was 13.8% (153/1111). Students' median score for knowledge was 3 out of maximum 6; attitude 3/4; awareness 4/6 and practice 1/2. Twenty-five students sustained a total of 28 NSI events in the past 12 months (proportion 16.6%, 95% confidence interval 10.9%-22.4%; incidence density 1.5/100 person-month); 13/25 did not report their NSI. Students in higher year of study were associated with higher rate of NSI (p = 0.02). Clinical and medical schools must address poor NSI-related knowledge, improve attitudes to reporting and provide instructions on safe handling of sharps. Surveillance of NSI specifically for medical students will provide important information to minimise harm.