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This was a cross-sectional study to start a cohort in two University Hospitals of two countries - Brazil and Colombia - for assessing the prevalence of needlestick and sharps injuries (NSI), the level of compliance with standard precautions (SPs), and knowledge on blood borne pathogens and associated factors among health students and professionals, within the framework of the implementation of the NR-32 standard. We created compliance scales based on 12 and 10 questions, for assessing knowledge. We used the Multinomial Poisson-Tweedie Regression to evaluate the relationship between knowledge and compliance with SPs within NSI. We evaluated 965 individuals (348 students and 614 professionals). The mean score points for level of knowledge was 10.98, with a median of 11 (10; 12) and α-Cr of 0,625. Compliance with SP had a mean of 30.74 and median of 31 (28; 34), with a α-Cr coefficient of 0.745, associated with country, group (student) and risk perception. Among the factors associated with the report of NSI, we singled out knowledge and compliance, country of origin, and full vaccination scheme against the Hepatitis B virus. We concluded that the level of knowledge and compliance were adequate among participants, but better among Brazilian participants, and it was associated with NSI reporting.