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Background This study developed a preliminary instrument to measure nurses’ infection prevention compliance against emerging respiratory infectious diseases and to verify the reliability and validity of the developed instrument. Method The participants were 199 nurses working at a university hospital with more than 800 beds and two long-term care hospitals. Data were collected in May 2022. Results The final version of the developed instrument consisted of six factors and 34 items, with an explanatory power of 61.68%. The six factors were equipment and environment management and education, hand hygiene and respiratory etiquette, infection risk assessment and flow management, protection of employees in contact with infected patients, ward access management of patients with infectious diseases, and wearing and removing personal protective equipment. We verified the convergent and discriminant validities of these factors. The instrument's internal consistency was adequate (Cronbach’s α=0.82), and the Cronbach’s α of each factor ranged from 0.71 to 0.91. Conclusion This instrument can be utilized to determine the level of nurses’ compliance with infection prevention activity against emerging respiratory infectious diseases and will contribute to measuring the effectiveness of future programs promoting infection-preventive activities.