Background: To identify nurses’ needs and problems in relation to research, and thereby enhance nursing research output, a valid measure of nurses’ attitudes toward research is necessary. Aim: This study aimed to develop, validate and conduct reliability analysis of a newly developed questionnaire assessing nurses’ attitudes towards conducting research. Subject and methods: We evaluated the content validity of the new questionnaire using an expert panel, while nurses with similar characteristics as the target group assessed for face validity. The content and face validity were determined by six experts and 11 nurses, respectively. The construct validity of the draft questionnaire was assessed using exploratory and confirmatory factor analysis from pilot-study data. The reliability (internal consistency and composite) of the final version of the questionnaire was then assessed. Statistical analysis was carried out using SPSS version 20 (IBM Corp., Armonk, NY) and SPSS AMOS version 18.0 for confirmatory factor analysis. Results: Two factors were extracted using an exploratory factor analysis. The confirmatory factor analysis confirmed that the two-factor structure, which contained 10 items in total, was the best fit to the data. The Cronbach’s alpha was good (0.81 and 0.77 for Factors 1 and 2, respectively). The composite reliability and discriminant validity of the final model were confirmed. Conclusion: We confirmed the validity and reliability of this questionnaire, making it a useful measure of attitudes towards conducting research among nurses.