文摘
To date there is no commercially-available serodiagnostic for women and men infected with Trichomonas vaginalis. Thirteen epitopes of the immunogenic T. vaginalis α-actinin (106.2-kDa) are detected by sera of women patients, and 5 epitopes, a subset of the 13, are detected by sera of men. A truncated recombinant protein called ACT-P2 (63.5-kDa) encoding the 5 epitopes is used for screening by ELISA for antibody in sera of women and men. Because ACT-P2 is poorly expressed in E. coli, we wanted alternative recombinant α-actinin proteins as serodiagnostic targets. We, therefore, constructed plasmids encoding two novel, small recombinant proteins of ∼25-kDa comprised of 15-mer peptides, each peptide of which contains one of the 13 epitopes. We refer to these novel proteins as α-actinin::string-of-epitopes1 (ACT::SOE1) and ACT::SOE2. We found the proteins to be unrecoverable from insoluble inclusion bodies without denaturing conditions, which rendered the proteins unsuitable for antibody detection. Thus, we synthesized a third ACT::SOE3 protein (72.4-kDa) with 7 epitopes that was synthesized in high amounts and was readily purified. Monoclonal antibodies to α-actinin detected ACT::SOE3 and ACT-P2 by ELISA. Further, we show that ACT::SOE3 is equal to ACT-P2 as a target protein for detection of serum IgG in positive sera of women and men. Data indicate that ACT::SOE3 is a target for screening of populations at-risk for this STI. Finally, the paper discusses the findings with regard to Point-of-Care diagnostic targets and vaccine candidates.
