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Abstract Background Variable and poor care quality are important causes of preventable patient harm. Many patients receive less than recommended care, but the extent of the problem remains largely unknown. The CareTrack Kids (CTK) research programme sought to address this evidence gap by developing a set of indicators to measure the quality of care for common paediatric conditions. In this study, we focus on one clinical area, ‘preventive care’ for pre-school aged children. Our objectives were two-fold: (i) develop and validate preventive care quality indicators and (ii) apply them in general medical practice to measure adherence. Methods Clinical experts (n = 6) developed indicator questions (IQs) from clinical practice guideline (CPG) recommendations using a multi-stage modified Delphi process, which were pilot tested in general practice. The medical records of Australian children (n = 976) from general practices (n = 80) in Queensland, New South Wales and South Australia identified as having a consultation for one of 17 CTK conditions of interest were retrospectively reviewed by trained paediatric nurses. Statistical analyses were performed to estimate percentage compliance and its 95% confidence intervals. Results IQs (n = 43) and eight care ‘bundles’ were developed and validated. Care was delivered in line with the IQs in 43.3% of eligible healthcare encounters (95% CI 30.5–56.7). The bundles of care with the highest compliance were ‘immunisation’ (80.1%, 95% CI 65.7–90.4), ‘anthropometric measurements’ (52.7%, 95% CI 35.6–69.4) and ‘nutrition assessments’ (38.5%, 95% CI 24.3–54.3), and lowest for ‘visual assessment’ (17.9%, 95% CI 8.2–31.9), ‘musculoskeletal examinations’ (24.4%, 95% CI 13.1–39.1) and ‘cardiovascular examinations’ (30.9%, 95% CI 12.3–55.5). Conclusions This study is the first known attempt to develop specific preventive care quality indicators and measure their delivery to Australian children in general practice. Our findings that preventive care is not reliably delivered to all Australian children and that there is substantial variation in adherence with the IQs provide a starting point for clinicians, researchers and policy makers when considering how the gap between recommended and actual care may be narrowed. The findings may also help inform the development of specific improvement interventions, incentives and national standards.