Urine free sialic acid (UFSA) is an important diagnostic biomarker for sialuria (GNE variants) and infantile sialic acid storage disease/Salla disease (SLC17A5 variants). Traditionally, UFSA has been measured using specific single-plex methodology in relatively small cohorts of patients with clinical symptoms suggestive ofthese disorders. The use of multiplex tandem mass spectrometry urine screening (UMSMS) has meant that UFSA can be measured semi-quantitatively in a muchlarger cohort of patients being investigated for suspected metabolic disorders. We hypothesised that theneuraminidase of Streptococcus pneumoniae may release free sialic acid from endogenous sialylated glycoconjugates and result inincreased UFSA levels. We conducted a retrospective review of clinical records of patients who were identified as having S.pneumoniae infection and who also had UMSMS at the timeof their acute infection. We identified three cases of increased UFSA detected by UMSMS screening that were secondary to S.pneumoniae sepsis. Additional testing ruled out geneticcauses of increased UFSA in the first patient. All three patients had overwhelming sepsis with multiorgan dysfunction which was fatal. Glycosylation abnormalities consistent with the removal of sialic acid were demonstrated in serum transferrin patterns in one patient. We have demonstrated in a retrospective cohort that elevation of UFSA levels have been observed incases of S.pneumoniae sepsis. This expands our knowledge of UFSA as a biomarker in human disease. This research demonstrates that infection with organisms with neuraminidase activity should be considered in patients with unexplained increases in UFSA.