A cross sectional questionnaire based study was undertaken to assess and compare knowledge, attitude and practice towards Human immunodeficiency virus (HIV) post exposure prophylaxis (PEP) amongst 400 participants including academicians, interns and students in two dental and medical colleges -Pune. Self-administered, structured, pretested, validated, close ended questionnaire was used. Unpaired t-test, analysis of variance (ANOVA), Tukey’s post hoc test, Z- test and chi square test for proportion were used. Off 400 questionnaires, 385 were returned giving a response rate of 96.25%. Significant proportion 342(88.8%) out of 385 participants, were found to have inadequate knowledge about PEP for HIV. Out of 55(4.2%) participants exposed to potentially infectious agents, eight initiated anti-retroviral therapy. Only three out of eight who initiated the therapy completed the course. Even though the attitude was favourable towards HIV PEP, adherence to guidelines for HIV PEP was poor which justify the need for a formal training and reinforcement.