Objective: The primary objective was to investigate temporal trends and between-practice variability of paediatric test use in primary care. Methods and analysis: This was a descriptive study of population-based data from primary care consultation records from January 1, 2007, to December 31, 2019. Children aged 0 to 15 who were registered to one of the 1,464 practices and had a diagnostic test code in their clinical record were included. The primary outcome measures were: 1) temporal changes in test rates measured by the average annual percent change (AAPC), stratified by test type, gender, age group, and deprivation level and 2) practice variability in test use, measured by the coefficient of variation (CoV). Results: 14,299,598 diagnostic tests were requested over 27.8 million child-years of observation for 2,542,101 children. Overall test use increased by 3.6%/year (95% CI 3.4 to 3.8%) from 399/1,000-child-years to 608/1,000 child-years, driven by increases in blood tests (8.0%/year, 95% CI 7.7 to 8.4), females aged 11-15 (4.0%/year, 95% CI 3.7 to 4.3), and the most socioeconomically deprived group (4.4%/year, 95% CI 4.1 to 4.8). Tests subject to the greatest temporal increases were fecal calprotectin, fractional exhaled nitric oxide (FeNO), and vitamin D. Tests classified as high use and high practice variability were iron studies, vitamin D, vitamin B12, folate, and coeliac testing. Conclusions: In this first nationwide study of paediatric test use in primary care, we observed significant temporal increases and practice variability in testing. This reflects inconsistency in practice and diagnosis rates, and a scarcity of evidence-based guidance. Increased test use generates more clinical activity with significant resource implications, but conversely may improve clinical outcomes. Future research should evaluate whether increased test use and variability is warranted by exploring test indications and test results, and directly examine how increased test use impacts on quality of care.