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Script concordance testing (SCT) is used to assess clinical decision-making. We explore the use of SCT to (1) quantify practice variations in infant lumbar puncture (LP) and (2) analyze physician's characteristics affecting LP decision making.Using standard SCT processes, a panel of pediatric subspecialty physicians constructed 15 infant LP case vignettes, each with 2 to 4 SCT questions (a total of 47). The vignettes were distributed to pediatric attending physicians and fellows at 10 hospitals within the INSPIRE Network. We determined both raw scores (tendency to perform LP) and SCT scores (agreement with the reference panel) as well as the variation with participant factors.Two hundred twenty-six respondents completed all 47 SCT questions. Pediatric emergency medicine physicians tended to select LP more frequently than did general pediatricians, with pediatric emergency medicine physicians showing significantly higher raw scores (20.2 ± 10.2) than general pediatricians (13 ± 15; 95% confidence interval for difference, 1, 13). Concordance with the reference panel varied among subspecialties and by the frequency with which practitioners perform LPs in their practices.Script concordance testing questions can be used as a tool to detect subspecialty practice variation. We are able to detect significant practice variation in the self-report of use of LP for infants among different pediatric subspecialties.