Objective. To examine the natural history, clinical manifestations, laboratory changes and outcomes of children with culture-negative osteomyelitis and to compare findings with cases of culture-positive infection treated during the same time period. Design. Retrospective review of consecutive admissions for osteomyelitis during the 4-year period 1998 through 2001, with a minimum follow-up of 1 year. Setting. A single urban children's hospital with a large orthopedic referral service. Clinical and laboratory measures. Age, gender, predisposing factors, clinical manifestations, maximum temperature, duration of pain, bone involved, laboratory changes, results of cultures from infected bone and blood and outcome after treatment. Results. A total of 85 patients fulfilled study criteria, of whom 40 were culture-negative. Compared with culture-positive cases, culture-negative osteomyelitis patients were less likely to have antecedent trauma (P = 0.0357) and overlying skin changes (P = 0.0001), duration of pain and other symptoms was longer (P = 0.0396) and skeletal residua were rare. They were also older, with this difference approaching statistical significance (P = 0.0586). Conclusions. Children with culture-negative osteomyelitis present initially differently from culture-positive cases and can be managed as presumed staphylococcal disease with excellent long term results.