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Background: The Democratic Republic of Congo (DRC) implemented a prevention of mother-to-child transmission (PMTCT) of HIV infection programme in maternal, newborn and child health (MNCH) services in 2001 with nurses as key personnel. To date there is no information in the DRC and specifically in Kinshasa with respect to compliance with PMTCT national guidelines., Aim: The study aimed at describing nurses’ compliance with the PMTCT national guidelines in selected PMTCT sites of Kinshasa., Methods: A descriptive cross-sectional study was conducted in Kinshasa with 76 nurses in 18 selected PMTCT sites. The nurses’ compliance with PMTCT national guidelines was assessed using a healthcare provider self-reporting questionnaire developed by the researchers., Results: The study showed that the mean score of nurses’ compliance with PMTCT national guidelines was 74% (95% CI: 69%-78%) which progressively decreased and was significantly different across different MNCH services (p = 0.025). With respect to categories of PMTCT recommendations, nurses were compliant with those related to education in labour and delivery, and antenatal services. Sociodemographic characteristics such as training, length of service and category of nurses did not influence nurses’ compliance score., Conclusion: These findings showed that nurses were noncompliant with PMTCT national guidelines, with the score level being 80% or more in the three MNCH services/units. Improvement of nurses’ ‘compliance with the PMTCT national guidelines requires effective monitoring of full integration of PMTCT as routine activities in MNCH care.