Background:Lyme disease is the most prevalent tick-borne illness in the United States, especially endemic in the Northeast and Upper Midwest. Distinguishing Lyme arthritis (LA), the most common manifestation of the disease in children, from septic arthritis (SA) can be challenging because of overlap in clinical presentations. This study examined the role of magnetic resonance imaging (MRI) as an adjunct to clinical and laboratory features used to differentiate between LA and SA in children and adolescents.Methods:The medical records and MRI scans of children who presented between 2009 and 2019 with an acute knee effusion ultimately diagnosed as LA or SA were retrospectively reviewed. Data collection included clinical information on the modified Kocher criteria (weight-bearing, fever, blood serology including white blood-cell [WBC] count, C-reactive protein [CRP], and erythrocyte sedimentation rate [ESR]), MRI findings, and serology confirmation of LA or bacterial SA. A total of 87 cases of confirmed LA and 9 cases of SA were identified.Results:The 2 cohorts had substantial clinical overlap with regard to the ability to bear weight, fever, and joint aspirate WBC count. Differences between the 2 groups in several MRI characteristics, specifically vastus lateralis myositis, subcutaneous edema, and lymphadenopathy, were significant. A multivariate analysis demonstrated that weight-bearing, CRP of <3 mg/L, absence of subcutaneous edema, myositis of multiple muscles including the vastus lateralis, and lymphadenopathy were predictive of LA.Conclusions:LA should be strongly suspected in endemic areas of the United States when children present with a knee effusion. The addition of MRI criteria to clinical and laboratory findings significantly improved the predictive value for identifying LA.Level of Evidence:Diagnostic Level III. See Instructions for Authors for a complete description of levels of evidence.