The aim of this study was to determine the frequency of autoantibodies among Jordanian patients with diabetes mellitus type 1. A total of 128 subjects were included with age range between 1year and 14 years, 45.3% (58) of which were female and 54.7% (70) were male. Samples were analysed for anti-Glutamic acid decarboxylase, anti-insulin, and anti-islet cell antibodies using mosaic pancreas (monkey)/cerebellum (monkey) instructions for the indirect immunofluorescence assay, Euroimmun, Germany. One hundred and twenty eight subjects with T1DM were enrolled in study; 58 female (45.3 %) and 70 male (54.7%). The overall frequency of any autoantibodies was 36.7% (47 of 128 subjects).Anti-GAD, anti- Islet cell and anti-insulin antibodies were detected in 30.46% (39 subjects), 25.78% (33 subjects), and 23.43% (30 subjects) respectively table-1. Anti-GAD and anti-islet cell antibodies were more frequent in male, and anti-insulin antibodies were more frequent in female's figure 1, 2, 3. Frequency of multiple antibodies more than one was observed in 38 patients (29.68%); whereas only one antibody (anti-insulin antibodies) was observed in 28 patients (21.87%). 18 of 19 subjects with positive anti-GAD antibodies had positive anti-islet cell antibodies. About one third of diabetes mellitus patients from Jordan had presence of autoantibodies such as anti-GAD, anti-islet cell, and anti-insulin in a single form or in a combination, which is suggest of having diabetes mellitus type 1. We recommend further large studies to be done in different areas of country.