Background: Spirometry is the most basic, widely used and effort-dependent pulmonary function test. It assesses the lung volumes and flows, and is ideally suited to describe the effects of restriction or obstruction on lung function. Therefore, keeping in view the clinical applications of spirometry, this study attempts to explore the knowledge and practice about spirometry among pediatricians. Materials and Methods: A questionnaire-based study was conducted across multiple centers in various hospitals in Riyadh, Saudi Arabia. The structured questionnaire, based upon knowledge and practice of spirometry, was distributed to 150 pediatricians in the various tertiary care hospitals in the metropolitan area of Riyadh. Results: Ninety-four percent of 113 pediatricians agreed that spirometry is a valuable tool in pediatric clinical practice. However, knowledge relating to spirometry was lacking among pediatricians, and about 86% of the study population did not demonstrate up-to-date knowledge of spirometry in pediatrics. Only 11% of pediatricians were very confident in interpreting spirometry results. No statistically significant association was observed between the distribution of responses relating to knowledge and practice of spirometry and the study variables including academic position, duration of practicing experience and number of patients attended daily. Conclusion: The results indicated that pediatricians in Riyadh were lacking adequate knowledge about the clinical applications of spirometry in their daily clinical practice. Hence, it was suggested that pediatricians should attend periodical training, workshops and continuous medical education programmes to enhance their knowledge. This should especially be performed during their pediatric residency training programmes, as spirometry is one of the essential components in clinical practice.