HPV (human papillomavirus) plays an important role in cervical cancer and may also play a role in vulvar cancer. TP53 mutation is common in a variety of cancers but its role in vulvar cancer is not well established. The aim of this study was to assess the prevalence of HPV infection and TP53 mutation as well as their correlation in vulvar cancer. Also, HPV detection and abnormal p53 expression were assessed in relation to age, co-existing vulvar intraepithelial neoplasia and vulvar dystrophy. Forty-eight samples of vulvar cancer were studied. DNA was extracted from formalin-fixed paraffin embedded tissue for polymerase chain reaction/Southern blot study with HPV 16 and 18 and L1 primers. Paraffin sections were immunostained (IHS) for p53 protein using three antibodies, p1801, CM1 and DO7. The p53 mutation was also screened using polymerase chain reaction (PCR) single-stranded conformation polymorphism (SSCP) and confirmed by sequencing. Overall, HPV was detected in 48% (23/48), of which 96% (22/23) were HPV 16 or 18. By IHS, p53 overexpression was detected in 46% of cases whilst TP53 mutations were identified in 21%. In HPV positive and negative tumours, p53 abnormal expression was detected in 39% and 52%, respectively, and TP53 mutation was found in 22% and 20%, respectively. Mutations were mainly found at codons 273 and 204. Age was not found to be associated with HPV detection. However, the presence of HPV (71%) or absence of abnormal p53 expression (65%) were higher in tumours with VIN3, but were not correlated with dystrophy.