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A descriptive exploratory research design was carried out to identify on Hospital Acquired Infection among Nurses in Western Regional Hospital, Pokhara. A total of 141 nursing personnel were selected by using census method. Self administered questionnaire were used for data collection. The obtained data were coded and entered in SPSS version 20 programs and analyzed by using descriptive, statistical methods such as frequency, percentage, mean and standard deviation. This study finding revealed that only 14.9 percent of the total respondent had average knowledge about hospital acquired infection. Comparing respondent’s level of knowledge according to their qualification, only 15.2 percent of graduate nurses had average knowledge and 14.5 percent of respondent had average knowledge who work in general ward whereas 15.2 percent had average knowledge who work in critical ward. Eighteen percent of the respondents with 6 months to 3 years work experience had average knowledge whereas above 5 years work experience had only 12.8 percent knowledge. It can be concluded that most of the nurses had below average knowledge on hospital acquired infection. Graduate nurses had more knowledge than undergraduate nurses but nurses with less than 5 years of working experience had more knowledge as compared to above 5 year experiences. So, there should need of providing in-service education to experienced nurses for up to date their knowledge on control of hospital acquired infection.