Escherichia coli and Staphylococcus epidermidis are predominant causes of neonatal sepsis, particularly affecting preterm infants. Susceptibility to infection has been attributed to ‘immature’ innate monocyte defences, but no studies have assessed global transcriptional responses of neonatal monocytes to these pathogens. We aimed to characterise the neonatal monocyte transcriptional responses to E. coli and S. epidermidis, and the role of common modifiers such as gestational age and exposure to chorioamnionitis, to better understand early life innate immune responses. RNA-sequencing was performed on purified cord blood monocytes from very preterm (<32 weeks gestational age, GA, n=11) and term infants (37-40 weeks GA, n=4) following standardised challenge with live S. epidermidis or E. coli. The major transcriptional changes induced by either pathogen were highly conserved between infant groups. We identified 994 genes specific to the E. coli response (E. coli signature), 1133 genes specific to the S. epidermidis response (S. epidermidis signature), and 8467 genes differentially expressed in response to both stimuli (common signature), highlighting a common extant neonatal monocyte response to infection. This common signature appeared to be largely mediated by TLR/NF-kB/TREM-1 signalling. Finally, we observed distinct transcription of a subset of genes between E. coli- and S. epidermidis-stimulated monocytes which appear to be differentially regulated by interferon-related signalling molecules. Our results demonstrate that neonatal monocyte transcriptional responses to E. coli and S. epidermidis (two structurally and biologically diverse neonatal sepsis pathogens) is largely shared, present early in gestation and not altered by prenatal exposure to chorioamnionitis. These data provide novel insights into the functionality of neonatal monocytes at birth, and highlight potential pathways that could be targeted to reduce the harmful effects of bacterial-induced inflammation in sepsis.