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Tuberculosis (TB) is an important cause of childhood death and morbidity worldwide. The diagnosis in the pediatric population remains challenging due to the paucibacillary nature of the disease. Intrathoracic lymphadenopathy is one of the most common manifestations of primary disease but is often difficult to sample. A retrospective case review was performed of children (younger than 16 years) suspected with intrathoracic TB lymphadenopathy who underwent an endobronchial ultrasound (EBUS)-transbronchial needle aspiration (TBNA) between January 2010 and 2020 in a London TB center. Ten children between 11 years 4 months and 15 years 9 months, with weights ranging from 48 to 95 kg, underwent EBUS-TBNA. All procedures were performed under conscious sedation with no reported complications. Six of 10 cases showed granulomas on rapid onsite histologic evaluation. Nine of 10 cases were confirmed to have Mycobacterium tuberculosis. Seven of 10 cases were culture positive with a mean turn-around time of 13.7 days; of these, 4 of 7 were smear positive. Six of 7 culture positive cases were also TB polymerase chain reaction (PCR) positive. TB PCR identified 2 further cases where microscopy and culture remained negative. One case had multidrug-resistant TB identified on TB PCR allowing early initiation of correct drug therapy. In our cohort, we show EBUS-TBNA is a safe and effective way of investigating intrathoracic TB lymphadenitis in children and a high diagnostic rate can be achieved. In high-resource settings, we should approach childhood TB with a standardized diagnostic approach and utilize EBUS-TBNA as a diagnostic modality. Samples should be sent for culture but also for molecular assays to timely identify TB and drug-resistant disease.