Dengue is an acute mosquito‐borne viral disease of humankind. Dengue fever, dengue haemorrhagic fever and dengue shock syndrome have become global public health problems in recent years. rDME‐G (recombinant dengue multiepitope protein that can specifically detect IgG) was produced in a 5‐litre fermenter in Escherichia coli for use in diagnosis. The culture was induced with 1 mM isopropyl β‐d‐thiogalactoside and cells were further grown for 4 h before harvesting. After fermentation, dry cell weight resulted in approx. 16.2 g/l. The rDME‐G protein was purified from inclusion bodies using affinity chromatography. The final yield of purified rDME‐G protein from fermentation resulted in approx. 168 mg/l of pure biologically active rDME‐G protein. The purity of rDME‐G protein was checked by SDS/PAGE analysis and the reactivity of this protein was further determined by Western blotting. The purified protein was used to develop an in‐house dipstick ELISA and tested using a panel of 60 patient sera characterized using the commercially available tests for detection of dengue antibody. We compared our results with IgG‐capture ELISA (Pan‐Bio, Windsor, QLD, Australia) and rapid IC (immuno‐chromatography) test (Pan‐Bio). By using rDME‐G protein as an antigen, in the dipstick ELISA, the results were in excellent agreement with commercial rapid IC test and IgG capture ELISA. These results show that the product has a promising potential to be used for diagnosis of dengue in both laboratory‐ and field‐based detection systems with minimum cost and a high degree of sensitivity and specificity.