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Introduction Pre‐treatment HIV drug resistance (PDR) is an increasing problem in sub‐Saharan Africa. Children are an especially vulnerable population to develop PDR given that paediatric second‐line treatment options are limited. Although monitoring of PDR is important, data on the paediatric prevalence in sub‐Saharan Africa and its consequences for treatment outcomes are scarce. We designed a prospective paediatric cohort study to document the prevalence of PDR and its effect on subsequent treatment failure in Nigeria, the country with the second highest number of HIV‐infected children in the world. Methods HIV‐1‐infected children ≤12 years, who had not been exposed to drugs for the prevention of mother‐to‐child transmission (PMTCT), were enrolled between 2012 and 2013, and followed up for 24 months in Lagos, Nigeria. Pre‐antiretroviral treatment (ART) population‐based pol genotypic testing and six‐monthly viral load (VL) testing were performed. Logistic regression analysis was used to assess the effect of PDR (World Health Organization (WHO) list for transmitted drug resistance) on subsequent treatment failure (two consecutive VL measurements >1000 cps/ml or death). Results Of the total 82 PMTCT‐naïve children, 13 (15.9%) had PDR. All 13 children harboured non‐nucleoside reverse transcriptase inhibitor (NNRTI) mutations, of whom seven also had nucleoside reverse transcriptase inhibitor resistance. After 24 months, 33% had experienced treatment failure. Treatment failure was associated with PDR and a higher log VL before treatment initiation (adjusted odds ratio (aOR) 7.53 (95%CI 1.61–35.15) and 2.85 (95%CI 1.04–7.78), respectively). Discussion PDR was present in one out of six Nigerian children. These high numbers corroborate with recent findings in other African countries. The presence of PDR was relevant as it was the strongest predictor of first‐line treatment failure. Conclusions Our findings stress the importance of implementing fully active regimens in children living with HIV. This includes the implementation of protease inhibitor (PI)‐based first‐line ART, as is recommended by the WHO for all HIV‐infected children