enhancing the detection of toxoplasma gondii via an anti-sag1 scfv-alkaline phosphatase immunoconjugate
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2019
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Abstract
The purpose of this study was to implement a fluorometric method for enhancing the detection sensitivity of Toxoplasma gondii in biological fluids. To address this challenge, we designed and produced a recombinant immunoconjugate tool based on a single-chain antibody fragment anti-T. gondii SAG1 antigen (scFvSG15) genetically fused to the bacterial alkaline phosphatase (AP) using 4-methyl-umbelliferyl-phosphate as fluorogenic substrate. The anti-SAG1 scFv-AP conjugate was fully bifunctional and was used successfully in different assays including immunoblot, fluorometric ELISA and direct immunofluorescence. The fluorometric immunoassay afforded an extremely low detection limit (1 tachyzoite/well), which was in agreement with the real-time PCR control test. The immunofluorescence imaging has provided captivating visual evidence of T. gondii detection. These results strongly suggest that the recombinant anti-SAG1-AP conjugate generated here might serve as useful and highly sensitive immunoassay probe to direct detect T. gondii in a one-step procedure, opening up new perspectives for diagnosis of toxoplasmosis. Keywords: Alkaline phosphatase, Fluorescent immunodetection, Recombinant conjugate, Single-chain antibody fragment variable (scFv), Toxoplasma gondii
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hannachi2019biotechnologyenhancing
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| Authors | ;Emna Hannachi;Aïda Bouratbine;Mohamed Mousli |
| Journal | PLoS genetics |
| Year | 2019 |
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