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Background: Vitamin B12 plays a role in hematopoiesis, neuronal functions, the synthesis of myelin and epithelium, and the mucosal functions of the gastrointestinal system, so patients with vitamin B12 deficiency may complain of a variety of symptoms. Objectives: This study aimed to investigate the association between vitamin B12 levels and clinical symptoms in children and adolescents. Methods: Patients visiting the pediatric outpatient clinic for any reason between April 1 and September 30, 2014 were enrolled in the study. Patients with active infectious diseases, with chronic diseases, or who had any disease or were taking any medications causing vitamin B12 deficiency were excluded from the study. The patient’s complaints were recorded. Each patient’s serum vitamin B12, 25-OH vitamin D, folic acid, and complete blood count were measured in addition to routine tests. Results: A total of 524 patients who were 8.3 ± 4.2 years old (2.6 months - 15.3 years), which consisted of 272 females (51.9%) and 252 (48.1%) males, met the inclusion criteria. The vitamin B12 level was less than 200 pg/mL in 166 (31.7%) of these patients and was less than 300 pg/dL in 302 (57.6%) of these patients. Vitamin B12 level was not associated with any complete blood count parameters except MCV. The patients with vitamin B12 deficiency were also 25-OH vitamin D deficient (P < 0.00). Symptoms, especially forgetfulness, depression, and anxiety were more common in the adolescent age group; these symptoms were significantly related to vitamin B12 deficiency (P < 0.05). Forgetfulness, fatigue, anxiety, and headache were the most common symptoms. There was no significant difference between the symptoms of the patients with vitamin B12 < 200 pg/mL and those with vitamin B12 < 300 pg/mL. Conclusions: With vitamin B12 deficiency defined as a vitamin B12 level < 300 pg/mL, we treated patients with vitamin B12 deficiency in order to prevent the progression of their symptoms to more serious complications. The early treatment of vitamin B12 deficiency may prevent the development of depression and anxiety in adolescents.