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为了研制一种可用于检测弓形虫的快速诊断胶体金免疫层析试纸条,本研究对弓形虫表面抗原(surface antigen,SAG)SAG1和SAG2的基因片段进行重构合成,与PET-28a(+)载体连接后,转化至大肠杆菌BL21(DE3)中表达,利用His亲和层析柱对融合蛋白进行纯化,获得SAG重组表位抗原,Western blot对重组蛋白免疫原性进行分析。用r SAG重组蛋白及r SPG分别划线,作为检测线和质控线,制作胶体金免疫层析试纸条,利用小鼠弓形虫阳性血清及小鼠阴性血清检验胶体金免疫层析试纸条检查效果。本研究成功表达纯化了SAG抗原表位的重组蛋白,且该重组蛋白具有较好的免疫原性。以此多抗原表位的重组蛋白作为检查抗原研制了胶体金免疫层析试纸条,能够快速检测弓形虫的阳性血清,为基层弓形虫病的快速诊断奠定了基础。
In order to develop a rapid diagnostic colloidal gold immunochromatographic strip for the detection of Toxoplasma gondii, this study reconstructed SAG1 and SAG2 gene fragments of Toxoplasma gondii, and compared with PET-28a +) Vector was transformed into E. coli BL21 (DE3) expression, the use of His affinity chromatography purification of the fusion protein to obtain SAG recombinant epitope antigen, Western blot analysis of recombinant protein immunogenicity. Using r SAG recombinant protein and r SPG were crossed, as the test line and the control line, the production of colloidal gold immunochromatographic strip, the use of Toxoplasma gondii positive serum and mouse negative serum test colloidal gold immunochromatographic test strip Article check the effect. In the present study, we successfully expressed the purified recombinant SAG antigen epitope protein, and the recombinant protein has good immunogenicity. The multi antigen epitopes recombinant protein as a test antigen developed colloidal gold immunochromatographic strip, can quickly detect Toxoplasma gondii positive serum, which laid the foundation for the rapid diagnosis of toxoplasmosis.