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According to WHO report (2008), In India it is reported that 50-60% of all neonatal death occur within the first month of life due to poor aseptic technique practicing in the labour room. Of these more than half may die during the first week of life, so study was aimed to assess the knowledge of staff nurses regarding aseptic techniques at selected hospital of Pilkhuwa. Materials and Methods: A descriptive survey design was adopted with 68 staff nurses selected by convenience sampling technique from selected hospital of Pilkhuwa, Hapur. Knowledge was assed using the structured knowledge questionnaire. The reliability coefficient was found 0.70 by Kudar Richardson-20 formula. Results: Findings of study showed that mean knowledge score of staff nurses was 13.13 with median score of 14. Standard deviation of knowledge score was 3.25. Maximum staff nurses i.e. 43 (63.24%) were having average knowledge whereas 25 (36.76%) having knowledge poor average, none of the staff nurse was having adequate knowledge regarding aseptic techniques. On computation of chi square, it was found that there was no significant association of the knowledge score of the staff nurse with any demographic variables at 0.05 level of significance. Conclusion: Study concluded that Staff nurses were not having adequate knowledge regarding aseptic techniques, so there is a need of proper educational programmes to enhance knowledge of staff nurses regarding aseptic techniques.