Purpose: The purpose of this study was to provide multifaceted interventions for nurses and to confirm changes in their knowledge, perception, and actual practice on the adequate storage, conditions, and transportation of culture specimens. Methods: A one-group pretest-posttest experimental design was conducted with 41 nurses in two general wards of a tertiary acute care hospital in Seoul. Multifaceted interventions including education, feedback, posting guidelines and reminders, and improvement in specimen management accessibility were provided from May 2019 to January 2020. Outcomes were measured before and after the interventions. Knowledge and perception of the nurses were evaluated using self-reported questionnaires and actual practice by observation. Results: After the interventions, the average knowledge score on transportation time was significantly increased (Z=-4.89, p<.001). However, the knowledge score on storage methods was not significantly increased. The perception score was significantly increased (t=-3.19, p=.003). The proportion of specimen storage times, places, and conditions managed properly was significantly increased from 43.0% (46/107) to 77.1% (84/109) (p<.001). The average transportation time of blood samples to the laboratory significantly decreased from 3 hours 36 minutes (±1 hour 52 minutes) to 3 hours 1 minute (±1 hour 41 minutes) (t=2.51, p=.013). The percentage of blood culture specimens arriving within 2 hours was increased significantly from 22.9% to 39.2% (χ2=6.90, p=.009). Conclusion: The interventions were effective. However, some specimens remained in the ward longer than expected after the interventions. This requires further interventions.