Sero-diagnostic evaluation of Toxoplasma gondii recombinant Rhoptry antigen 8 expressed in E-coli

Background: Toxoplasma gondii infects all warm-blooded animals, including humans. Early diagnosis and determining the infective stage are critical for effectively treating immunosuppressed individuals and pregnant women with toxoplasmosis. Among the rhoptry proteins of the parasite, Rhoptry protein 8 (ROP8), is known to be expressed during the early stages of T. gondii infection and is involved in parasitophorous vacuole formation. In this study, we have investigated the diagnostic efficacy of recombinant ROP8 (rROP8). Methods: The ROP8 gene was cloned into pCOLD I DNA vector and expressed as a soluble recombinant antigen in Escherichia coli. Expressed ROP8 protein was evaluated using western blot method. Results: Western blot analysis of purified rROP8 antigen using 200 T. gondii-infected human serum samples, as well as non-infected serum controls, allowed for the successful identification of toxoplasmosis-positives, yielding a 90% sensitivity and 94% specificity. Conclusion: Our findings indicated that rROP8 antigen expressed in E. coli was able to detect toxoplasmosis in infected human serum with specificity and sensitivity suggesting that rROP8 antigen represents a valid diagnostic marker for toxoplasmosis.