Immunodiagnosis and molecular validation of Toxoplasmagondii-recombinant dense granular (GRA) 7 protein for the detectionof toxoplasmosis in patients with cancer

Arab-Mazar, Zahra and Fallahi, Shirzad and Koochaki, Ameneh and Haghighi, Ali and Seyyed Tabaei, Seyyed Javad (2016) Immunodiagnosis and molecular validation of Toxoplasmagondii-recombinant dense granular (GRA) 7 protein for the detectionof toxoplasmosis in patients with cancer. Microbiological Research, 183. pp. 53-59.

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Abstract

tSerological assays for the diagnosis of toxoplasmosis mostly rely on the tachyzoite specific antigens of Tox-oplasma gondii, which are difficult to produce by conventional methods. The aim of this study was to cloneand express of GRA7 protein of T. gondii and evaluate its potential for immunodiagnosis of toxoplasmosisin cancer patients. As well as validate the results using a new molecular assay, LAMP technique. The GRA7gene was successfully cloned, expressed and purified by affinity chromatography and the production wasevaluated by SDS PAGE, dot blot and western blot analyses. The rGRA7 was used for developing an ELISAbased on the rGRA7 using sera from patients with toxoplasmosis and healthy controls. Furthermore, 50serum samples from leukemic children infected with toxoplasmosis and 50 seronegative controls wereincluded to evaluate the sensitivity and specificity of rGRA7 based ELISA. Finally, the LAMP technique wasused to assess the accuracy and validity of the results obtained by rGRA7 based ELISA. The consistency ofthe results of two tests was determined by using the Kappa coefficient of agreement. The rGRA7 showedhigher and optimum immunoreactivity with 1:100 dilution of serum from Toxoplasma infected patients.The sensitivity and specificity of test were calculated as 92 and 94%, respectively. According to the Kappacoefficient of agreement, there was a significant conformance between the results obtained by ELISAbased on the rGRA7 and the results of LAMP technique (≈96%, P < 0.001). Findings of the present studyshowed that rGRA7 can be used as a potential immunogenic antigen for developing immunodiagnostictools for immunodiagnosis of toxoplasmosis in patients including patients with cancer.

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