Background Prescription-checking is becoming increasingly popular in medical institutions, but the prescription-checking ability of pharmacists needs improvement. This study aimed to explore the key aspects of the prescription-checking training and provide an empirical reference for standardizing the prescription-checking training for pharmacists in medical institutions.Methods Trainees who were willing to complete the Likert questionnaire were enrolled in this study. Percentage and composition ratio were used for statistical description. The chi-square test and exploratory factor analysis were used for statistical analysis. A difference with p<0.05 was considered statistically significant.Results The questionnaire had good internal consistency and validity. The overall satisfaction of the trainees was 90%. Three dimensions, training organization, teaching mode as well as consolidation and assessment of knowledge were extracted using exploratory factor analysis. The average examination score of the 20 courses was 89.21±2.62 points. From the perspective of needs, 94.66% of the trainees preferred face-to-face lectures, 89.33% of the trainees expected high professional skills of the lecturers, and 62.27% of the trainees believed that clinical expertise was highly desirable.Conclusion There was a great demand for prescription-checking training among pharmacists. Trainees enrolled in this training showed high satisfaction and good examination results. The key aspects of prescription-checking training were training organization, as well as consolidation and assessment of knowledge. It is recommended to conduct training in a stratified manner according to pharmacists’ educational background, in order to effectively improve their prescription-checking competence. The pharmacists preferred face-to-face lectures and interaction to supplement clinical disease-related knowledge.