Introduction: Hand hygiene is most important measure to minimize the ratio of transmission of health care associated infections (HCAIs) as this is one of the major problem especially in developing countries and most of the time caused behind it is poor hand hygiene of health care providers especially nurses as they have maximum interaction to the patients .This study was conducted to assess the knowledge and practices regarding hand hygiene among the nurses of DHQ hospital Okara. Methodology: A self-structured questionnaire with the help of WHO guidelines base cross sectional study was done on the sample of 100 nurses among the nurses of DHQ hospital Okara. Likert scale was used to assess their knowledge and they knowledge were scored as good (13-15), average (9-12) and poor (0-8) according to their responses. Practices were also assessed by questionnaire. Results: Most of the participants showed having good (51%) and average (39%) knowledge and there are only some nurses (4%) who showed poor results. But despite of having sufficient knowledge there was no implementation of hand hygiene because of lacking of facilities, over burden of patients, shortage of time. Conclusion: To improve the practices of hand hygiene continues training sessions, sufficient nurses, and provision of all facilities needed for the hand hygiene are most important things.