Background: Tuberculosis (TB) is one of the major health challenges in many developing countries and in Sudan in particular. The aim of this study was to screen pediatric patients suspected with lymph Node (LN) TB for the presence of tuberculosis. Methodology: This study investigated retrospectively 42 LN biopsies taken from pediatric patients for evidences indicating the presence of Mycobacterium Tuberculosis (MT). Hematoxylin and Eosin (HE), Zielnelson (ZN), Immunohistochemistry (IHC) and Polymerase Chain Reaction (PCR) techniques were used for diagnosis. Results: For HE giant cell granuloma and caseation were evidenced in 33/42(78.6%), since epitheloid granuloma was evidenced in 9/42(21.4%). Positive ZN, IHC and PCR were indicated in 1/42(2.4%), 33/42 (78.6%) and 33/42(78.6%), respectively. Conclusion: pediatric TB is still prevalent in some parts of Sudan. More effective health strategies are urgently needed in Sudan, particularly in eastern Sudan to control the disease.