Abstract Background: Hearing impairment is a partial or total inability to hear, may occur in one or both ears. In children hearing problems can affect the ability to learn spoken language and result in loneliness. Aim of the study: assess knowledge and attitude of nurses regarding Newborn Hearing Impairment and screening at Health Centers and clinics in Assiut City. Subjects and Methods: Descriptive cross-sectional design was used. A Total coverage sample of 235 nurses has participated. Tool 1: A structured self-administered questionnaire, it included three parts, Part one: personal data, Part two: questions related to attending training course regarding newborn hearing screening, Part three: knowledge about Newborn Hearing Impairment (NHI) and screening. Tool 2: It included two scales, Part one: Attitude assessment scale about NHI, and Part two: Attitude assessment scale about Newborn Hearing Screening. Results: 34.5% of nurses had poor knowledge, and 69.4% of them had a positive attitude regarding NHI, while 68.9% had a negative attitude against newborn hearing screening. There is a statistically significant difference between the score of nurses’ knowledge and their age. Conclusion: Deficit in nurses’ knowledge regarding newborn hearing impairment and screening; and fluctuation in their attitude between negative and positive. Also, there were statistically significant differences between the level of nurses’ attitude about newborn hearing screening, their age, and years of experience. Recommendations: in-service training programs should be developed and implemented for nurses