Giardia duodenalis is considered the most common cause of parasitic diarrhea worldwide. Genetic studies revealed that at least eight assemblages (A-H) exist. Of these assemblages, A and B are found primarily in human beings and occasionally in animals. The association between clinical symptoms and G. duodenalis assemblages is controversial. The aim of the present study was to determine the assemblages of G. duodenalis prevalent among Egyptian children with diarrhea. Therefore, 96 positive stool samples for Giardia by light microscopy were subjected to multilocus genotyping targeting the triose phosphate isomerase (tpi), β-giardin (bg), and glutamate dehydrogenase (gdh) genes. Amplified polymerase chain reaction (PCR) products were then purified, sequenced, and aligned with reference strains to determine the assemblages of the Giardia isolates. Out of the 96 microscopically positive stool samples for Giardia, 77 (80 %) were successfully amplified and sequenced at least at one locus. Of these, 21 (27.3 %) were shown to be assemblage A, 54 (70.1 %) assemblage B, while discordant sequence typing results were observed in 2 (2.6 %) samples. AII was the predominant subassemblage of assemblage A, while it was generally difficult to further classify assemblage B. It was concluded that infection with assemblage B was more common than that with assemblage A. No associations between epidemiological information and assemblage were detected, except with age. Although infections with assemblage B were more frequently associated with abdominal pain and acute diarrhea than with assemblage A, the difference was not statistically significant.