In the present study, there were two groups of diabetic patients. The first group was newly diagnosed diabetic patients of less than one year duration of disease. They were 40 patients. The second group was diabetic patients with more than one year duration of disease. They were 30 patients. The control group was 30 normal healthy children. Evidence of virus infection was detected by tissue culture isolation, neutralization test, RT-PCR, IgM, and IgG specific antibodies for coxsackie B viruses. There was significant increase in percent of tissue culture isolation of EV in group I more than group II while the percent in the two groups were significantly increased than the control group. Identification of the type of EV by neutralization test revealed that most of the type cases were Coxsackie B4 virus and one case was Coxsackie B6 virus (the most important diabetogenic strain). Viral RNA detection by PCR was done and revealed that most of cases in group I diabetic patients were positive for enterovirus while one case in group II of diabetic patients was positive. As regards Coxsackie virus B IgM antibodies positivity, there was increase in the percent in group I than group II, as it is a marker of acute infection. As regards Coxsackie virus B IgG antibodies positivity, there was no significant difference between group I and group II of diabetic patients, as it is a marker of past infection and it persists for years after the first episode.