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Objectives: To investigate the association between tonsillitis and the risk of newly diagnosed ankylosing spondylitis (AS). Methods: We used 2003–2012 data from Taiwan’s National Health Insurance Research Database to conduct this nationwide, population-based, case-control study. We identified AS patients newly diagnosed between 2005 to 2012 as the study group and selected age, sex and index-year matched (1:6) non-AS individuals as controls. The association between tonsillitis and risk of newly diagnosed AS was determined by calculating odds ratios (ORs) with 95% confidence intervals (CIs) using conditional logistic regression analysis. Results: We identified 37,002 newly diagnosed AS cases and 222,012 matched non-AS controls. Patients with AS were more likely to have tonsillitis (aOR 1.46, 95% CI 1.43–1.50), appendicitis (aOR 1.29, 95% CI 1.13–1.48) and periodontitis (aOR 1.35, 95% CI 1.31–1.38) than non-AS control subjects. The association between tonsillitis and AS was consistent using varying definitions for tonsillitis, and we further found that a high frequency of visits for tonsillitis, a high medical cost for tonsillitis and a long interval between diagnosis were associated with newly diagnosed AS in a dose-response manner. Furthermore, the association between tonsillitis and AS appeared to be stronger in females (aOR 1.59, 95% CI 1.53–1.65) than those in males (aOR 1.39, 95% CI 1.35–1.44). Conclusions: The present study revealed an association between AS risk and prior tonsillitis and indicates the need for vigilance of AS-associated symptoms in patients who had been diagnosed with tonsillitis, particularly in females. [ABSTRACT FROM AUTHOR]