Immunodiagnosis and molecular validation of Toxoplasma gondii‐recombinant dense granular (GRA) 5 protein for the detection of toxoplasmosis in hemodialysis patients

Abstract

Toxoplasmosis causes serious complications in immunocompromised and pregnant women. Serological tests for the detection of toxoplasmosis are often designed from parasitic tachyzoites antigens. The process of producing these antigens is very difficult. The purpose of this study was evaluation of T. gondii‐rGRA5 for the immunodiagnosis and molecular detection of Toxoplasma infection using enzyme‐linked immunosorbent assay (ELISA) and LAMP methods in hemodialysis patients. The GRA5 gene was successfully expressed and purified by affinity chromatography assay and evaluated by western blot. Then it was used to design an ELISA assay. A total of 260 samples were tested for anti‐Toxoplasma IgG and IgM antibodies using a commercial ELISA kit and designed ELISA kit. Finally, the LAMP method was used to evaluate the precision and reliability of the results obtained by commercial and designed ELISA kits. The consistency of the results of two methods was analyzed using the Kappa coefficient of agreement. The rGRA5 revealed higher immunoreactivity with 1:100 dilution of sera from toxoplasmosis patients. The specificity and sensitivity of the assay were 93% and 96%, respectively. According to the Kappa coefficient, there was a substantial correlation between the results of ELISA and LAMP based on rGRA5 (≈98%, p < 0.001). Also it showed that rGRA5 protein can be used as an antigenic protein for designing sero‐diagnostic tests to identify Toxoplasma infection especially in hemodialysis patients.

Mining the Proteome of Toxoplasma Parasites Seeking Vaccine and Diagnostic Candidates

“…The specificity and sensitivity of enzyme-linked immunosorbent assay (ELISA) were 93% and 96%, respectively. The loop-mediated isothermal amplification (LAMP) method also corroborated the accuracy and reliability of the results obtained by designed and commercial ELISA kits [78].…”

Development and evaluation of indirect enzyme-linked immunosorbent assay using recombinant dense granule antigen 7 protein for the detection of Toxoplasma gondii infection in cats in Thailand

“…Consequently, they have been considered as a good serodiagnostic test for the detection of T. gondii antibodies both in animals and humans [21]. Many recombinant antigens have been previously studied, namely, GRA1 [21][22][23][24], GRA2 [25][26][27], GRA4 [11], GRA5 [19,28], GRA6 [26,29], GRA7 [18,[21][22][23][24]26,[30][31][32][33], GRA8 [34,35], and GRA15 [26]. GRA7 provides a very strong humoral immune response in the acute stage of infection [3] and is also recognized as a good serological marker for anti-T. gondii detection in the chronic stage [23,31].…”