BackgroundTyphoid fever, an infective bacterial disease, is capable of causing fatal systemic infection in humans, and in an era of antimicrobial resistance, it has become of public health importance. This study aimed to investigate the laboratory diagnosis of Salmonella bloodstream infection, its serotype, antimicrobial resistance pattern, and seasonal variation at a tertiary care children’s hospital.MethodsWe undertook a retrospective, cross-sectional study by reviewing hospital-based laboratory records of patients whose blood culture samples were submitted from the outpatient department to the laboratory of a tertiary care children’s hospital in Kathmandu, Nepal, from January 2017 to January 2019.ResultsAmong the total blood culture samples obtained (n = 39,771), bacterial isolates (n = 1,055, 2.65%) belonged either to the Genus Enterobacteriaceae or Genus Acinetobacter. Altogether (n = 91, 8.63%), isolates were positive for Salmonella spp., which were further identified as Salmonella enterica subsp. enterica ser. Typhi (n = 79, 7.49%), Salmonella enterica subsp. enterica ser. Paratyphi A (n = 11, 1.04%), and Salmonella enterica subsp. enterica ser. Paratyphi B (n = 1, 0.1%). The median age of patients was 6 years (IQR: 4–9), with male and female patients constituting (n = 53, 58.24%; OR, 1.0; 95% CI, 0.60–1.67) and (n = 38, 41.76%; OR, 0.98; 95% CI, 0.49–2.05) cases, respectively. The disease was observed throughout the year, with a high prevalence toward the spring season (March–May). An antibiogram showed resistance more toward nalidixic acid with S. Typhi, comprising half the isolates (n = 52, 65.82%; p = 0.11). Resistance toward β-lactams with β-lactamase inhibitors (amoxicillin/clavulanate; 1.27%) was seen in a single isolate of S. Typhi. The multidrug resistance pattern was not pronounced. The multiple antibiotic resistance (MAR) index was in the range between 0.14 and 0.22 in S. Typhi and 0.22 and 0.23 in S. Paratyphi.ConclusionSalmonella Typhi was the predominant ser. Infection was common among children between 1 and 5 years of age, showing male predominance and with the spring season contributing to a fairly higher number of cases. Antimicrobial susceptibility testing of S. Typhi showed more resistance toward nalidixic acid, with only a single isolate resistant to β-lactamase inhibitors (amoxicillin/clavulanate). Alarming multidrug resistance patterns were not observed. The MAR index in this study indicates the importance of the judicious use of antimicrobials and hospital infection prevention and control practices.