Objective: To explore the effect of comprehensive nursing intervention on the lactation effect of mothers with maternal separation through qualitative research. Methods: From March to September 2019, 13 women in our department who required maternal separation were included in this study and given comprehensive nursing intervention. The general data of primiparas were obtained by questionnaire. Postpartum women were interviewed by semi-structured interview and the interview data were analyzed by the seven-step analysis method of phenomenology. Results: These postpartum women with maternal separation lacked knowledge and skills of lactation, experienced the difficulty of lactation and confusion of the quality of breast milk. After the comprehensive nursing intervention, they enhanced their knowledge and skills of lactation. Therefore, the effect of lactation is improved, and the quality of breast milk is maintained or improved. The evaluation of postpartum women to comprehensive nursing is high. Conclusion: Comprehensive nursing intervention has a positive effect on lactation and is worth popularizing and applying in clinic.