Human rotavirus is a major etiologic agent for infantile diarrhea worldwide. It is responsible for up to 3.3 million deaths per year in children in developing countries. Various rapid and sensitive techniques have been developed to readily diagnose rotavirus gastroenteritis. In the present study, we compared the sensitivity and specificity of immunochromatography and RNA-polyacrylamide-gel electrophoresis (PAGE) methods with enzyme immunoassay (EIA) for diagnosis of group A rotavirus infection in 200 stool samples from children younger than 5 years old with acute gastroenteritis. Rotavirus was detected in 57 (28.5%) samples by EIA, 52 (26%) samples by ICG and 52 (26%) samples by RNA-PAGE. There was no significant difference between the three methods (P=0.8) nor between EIA and ICG (P=0.57) and EIA and RNA-PAGE (P=0.57). Furthermore, in comparing these methods with age variables, the present study found that the sensitivity and specificity of ICG and RNA-PAGE compared with EIA were 87.7%, 98.6% and 91.2%, and 100%, respectively (P>0.05). Results of the present study demonstrate that the sensitivity and specificity rates for ICG and RNA-PAGE were as high as EIA. It seems that all the three methods are reliable and suitable for detection of group A rotavirus infection in children affected by enteric diseases.