ObjectiveTo construct a good reliability and validity of questionnaire for patients participating in hand hygiene (HH) promotion in medical institutions. MethodsQuestionnaires for patients participating in HH promotion in medical institutions were constructed by research group, 195 hospitalized patients and 207 health care workers(HCWs) in two medical institutions of Xi’an city were randomly selected and investigated, questionnaire items were analyzed with critical ratio, correlation coefficient, and factor analysis method. ResultsOriginal questionnaires included questionnaires for patients and HCWs, each had 3 dimensions and 20 items. The revised formal questionnaires for patients and HCWs had 17 items respectively, questionnaires for patients included 8 items in cognitive dimension, 7 items in willingness dimension, all were closed problems; 2 items for participation ways, both were openended questions. Questionnaires for HCWs included 9 items in cognitive dimension, 5 items in willingness dimension, 1 item for participation status, all were closed problems; 2 items for participation ways, both were openended questions.ConclusionThe questionnaires for patients participating in HH promotion in medical institution are further optimized though analyzing and revising the items based the investigated data.