Background: Globally every year, 2.7million neonates die during the neonatal period. In Ethiopia Neonatal, mortality is higher among many African region and other undeveloped countries. Recognition and early sought of medical care towards neonatal danger sign has many implications for decreasing neonatal morbidity and mortality. Despite the importance of, early sought of medical care towards neonatal danger sign the practice is low in Ethiopia. Objective: To assess knowledge, attitude and practice on neonatal danger sign and associated factors among PNC attendant mothers in Dessie referral Hospital, North East Ethiopia. Method: A Facility-based cross-sectional study employed. The data collected from 340 mothers from March to April 2018 by consecutive sampling technique. The data collection tool was adapted from WHO, Neonatal Health and it was collected by using Interview based structured questionnaire. Descriptive statistics, binary and multivariable logistic regression analysis employed to identify the factors associated with Newborn danger sign practices. Variables with a p-value < 0.05 identified as statistically significant factors. Result: The proportion of good newborn danger sign practice was 220 (92.8%) in Dessie Referral Hospital. 58(17.1%) of respondents were knowledgeable and 146(42.9%) of mothers had favorable/positive attitude towards newborn danger sign. Mother’s income between >1500 Birr/month 8.194(AOR: 95% CI: 2.583, 25.995), Mothers who live in rural side4.210 (AOR: 95% CI: 1.41, 12.576), Mothers who has instrumental delivery 9.843 (AOR: 95% CI: 5.148, 30. 31) were positive predictors for newborn danger sign practice. Conclusion and Recommendation: The magnitude of newborn danger sign practice in these specific setting found to be good but the mothers knowledge and attitude towards newborn danger sign is poor. Promotion of intensive education program on at all level, integrating it with post-natal and child health service and focusing on urban and low socioeconomic status mothers is recommended.