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<i>Background and objectives</i>: Stress in nursing students is a very common experience, especially when they face clinical practice. The aims of this study were to perform a transcultural adaptation and to examine the reliability and repeatability of the Instrument for the Assessment of Stress in Nursing Students for a Spanish population. <i>Methods</i>: A test&#8722;retest analysis was carried out in two face-to-face sessions with the students with a lapse of 10 days between the two sessions. A cross-sectional descriptive study was carried out between the months of May and June 2018. Sixty-four nursing students were recruited with a consecutive sampling method that targeted individuals in the freshman class. <i>Results</i>: A good internal consistency was shown for the total score (&#945; = 0.8861) and for each of the six domains. The test&#8722;retest reliability, using the Wilcoxon paired test, was not significant, indicating no differences between the total scores or the domain scores (<i>p</i> &#8805; 0.05). Finally, Bland and Altman plots of visual distributions did not show differences between the total scores and the domain scores. <i>Conclusion</i>: The Instrument for the Assessment of Stress in Nursing Students was shown to be a reliable tool for measuring stress factors among Spanish nursing students.