Abstract: Objective To prepare a monoclonal antibodies (mAbs) against glutamate dehydrogenase (GDH) of Plasmodium falciparum (FCC1/HN strain) and establish colloidal gold-immunochromatographic assay (GICA) for diagnosis of Plasmodium falciparum malaria. Methods Recombinant GDH was used to immunize Balb/C mice and the mAbs against GDH were prepared using hybridoma technique followed by identification of IgG isotype and its affinity. Protein-G affinity chromato- graphy was employed to purify the antibodies, which were labeled with colloidal gold for establishment of GICA for Plasmodium falciparum detection. Results Six mAbs were obtained and identified as IgG₁(κ) of IgG isotypes with affinity constants (Kaff) ranging from 1×10^-8 to 2.8×10^-10. GICA had a sensitivity of 86.66% and specificity of 96.43% for Plasmodium falciparum detection compared with routine microscopic examination. Conclusion The established GICA is rapid and accurate for Plasmodium falciparum detection with such potential utility as for instant diagnosis of Plasmodium falciparum malaria.