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Background: Motivational interviewing (M.I.) is an option for modifying an individual′s behavior. It is used as an educational method in recent years. The aim of our study was to indicate whether or not education, using lecture alone and lecture with M.I., would affect the performance of nursing personnel regarding their hand hygiene. If so, which of these two methods were most effective for this purpose? Methods: This was an interventional study conducted in Bahrami Pediatric Hospital in Tehran. The study population consisted of all nursing personnel in the hospital. Considering the responsibilities of different nurses and their educational status, we divided them into three classes. The participants of each class were randomly assigned to either the control or experiment groups training lecture alone or lecture with M.I., respectively. We used Independent-t, Paired-t, Mann-Whitney U and Wilcoxon signed ranks tests for analysis. Results: Education using lecture alone improved the hand hygiene performance of nursing personnel only in the first and third classes ( P = 0.002 and P = 0.001, respectively). Similarly, lecture combined with M.I. improved the hand hygiene performance of personnel in the first and third classes ( P < 0.001 and P = 0.004, respectively). The latter method was more effective compared to lecture alone in the first and third classes ( P < 0.001 and P = 0.013, respectively). Conclusions: Education based on lecturing improves hand hygiene performance among nursing personnel. It will be more effective if combined with M.I.