Introduction: Determination of the efficacy of crude Entamoeba histolytica extract and its fractions (FI,FII & FIII) in antiamoebic antibody detection by ELISA. Material and Methods: Entamoeba histolytica NIH:200 was cultured axenically and different fractionsof its crude amoebic extract, i.e. FI, FII & FIII, were obtained by column chromatography. Efficacy of Crudeand fractioned antigens for antiamoebic detection by ELISA was compared in acute amoebiasis cases. For thispurpose, a total of 25 cases including 15 amoebic liver abscess cases, 10 acute amoebic dysentery and 10control cases were included into the study. Results: Crude Entamoeba histolytica extract and fraction I are more useful for antiamoebic antibodydetection. While, efficacy of FII & FIII is lower and theses two extracts are not as effective in antiamoebicantibody detection. By using these antigens the sensitivity of ELISA reduces while using FI for ELISA hasquite a high sensitivity. Conclusion: In intestinal and extraintestinal invasive amoebiasis humoral immune response isaccompanied by antibody protection against amoebic antigens. Antibody present in sera of study groups weredetected by employing ELISA and it was found that ELISA, especially by using fraction I of Entamoebahistolytica extract, is 90_100% specific in detecting antiamoebic antibody.