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Objectives This study aimed to identify potential categories of rotation stress among nurses undergoing standardised training and to explore the relevant factors associated with each profile.Design Cross-sectional study.Setting Data were collected in November 2024 from three hospitals in Zunyi City, Guizhou Province, China.Participants Nurses undergoing standardised training were recruited for this study.Methods Convenience sampling method was used to recruit standardised training nurses in November 2024 from three hospitals in Zunyi City, Guizhou Province. The survey instruments used included demographic characteristics questionnaire, the Nursing Job Rotation Stress Scale and the Maslach Burnout Inventory. Latent profile analysis method was used to analyse rotation stress characteristics of nurses during standardised training. Additionally, logistic regression was performed to identify the factors influencing different characteristics.Results A total of 493 nurses completed the questionnaires, of which 453 were valid, resulting in a validity rate of 91.88%. Rotation stress was classified into two profiles: ‘Low Emotional Response–Stress Adaptation Group’ (21.5%) and ‘High Emotional Response–Stress Distress Group’ (78.5%). Univariate analysis showed that highest degree (χ 2=11.389, p=0.001), monthly night shifts (χ2=33.913, p<0.001), employment type (χ2=20.858, p<0.001), new graduate (χ2=12.319, p<0.001), pretraining work experience (χ2=35.754, p<0.001) and training duration (χ2=15.357, p=0.002) significantly influenced the two subgroups. Multivariable regression analysis revealed significant associations of monthly night shifts, pretraining work experience, training duration and burnout level (p<0.05).Conclusions Nurses undergoing standardised training exhibit two distinct rotation stress profiles. Monthly night shifts, pretraining work experience, training duration and burnout are significant factors. Nursing managers should implement targeted interventions such as mindfulness, laughter therapy and emotional freedom techniques to mitigate stress and thereby enhance the quality of standardised training.