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用昆虫细胞中表达的弓形虫主要表面抗原（SAGI）为试剂检测弓形虫IgG抗体。方法将昆虫细胞中表达的重组蛋白（rSAGI）通过单克隆抗体制备的亲和层析柱纯化．用纯化的rSAGI为抗原试剂检测弓形虫病血清，与其他试剂盒检测结果比较，然后再用免疫印迹试验确认．结果重组杆状病毒Bac304感染的Sf9细胞与空载bacmid感染的细胞比较，Bac304的SDS裂解物比空载株在SDS—PAGE考马斯亮蓝染色胶上明显多出一条带，分子量约为30kDa．在恒定条件下，分批收集Bac304感染的昆虫细胞，用免疫荧光鉴定，95％以上的细胞可被感染．在Balb／C小鼠中生产的抗SAGI的单抗XllA10C与Bac304的表达产物发生反应，用纯化的单抗与CNBr—Sepharose4B偶联制备的亲和层析柱从1．3X109感染Bac304的昆虫细胞裂解液中获得1．4mgrSAGI，以纯化rSAGI为抗原，采用EIA法检测了412份妇产科就诊者及不明原因淋巴结肿大等病人血清，检出弓形虫lgG抗体阳性13例，其中10例经全虫体蛋白印迹法确认为阳性．结论昆虫细胞表达的重组SAG1能识别人弓形虫病血清，可以用作诊断试剂．
Toxoplasma gondii IgG antibodies were detected using the Toxoplasma major surface antigen (SAGI) expressed in insect cells. Methods Recombinant protein (rSAGI) expressed in insect cells was purified by affinity chromatography on monoclonal antibody. Toxoplasmosis serum was detected using purified rSAGI as an antigen reagent, compared with the results of other kits, and then confirmed by Western blotting. Results Bacillus subtilis Bac304 infected Sf9 cells compared with empty bacmid infected cells, Bac304 SDS lysate than the empty vector SDS-PAGE Coomassie brilliant blue staining gel significantly more than a band, molecular weight of about 30kDa. Bac304-infected insect cells were collected in batches under constant conditions and more than 95% of the cells were infected by immunofluorescence. The anti-SAGI monoclonal antibody XllA10C, produced in Balb / C mice, reacts with the expression product of Bac304 and was infected with 1.3X109 insect cells infected with Bac304 using an affinity column prepared by coupling the purified monoclonal antibody with CNBr-Sepharose 4B 1.4 mg rSAGI was obtained from the lysate and rSAGI was purified as antigens. The serum of 412 obstetrics and gynecology patients and patients with unexplained lymph node enlargement were detected by EIA method. Toxoplasma lgG was detected in 13 of 10 patients All the worms were confirmed as positive by western blot. Conclusion Recombinant SAG1 expressed by insect cells can recognize human Toxoplasma gondii serum and can be used as a diagnostic reagent.