间接免疫酶染色检测小鼠组织内弓形虫及抗原

Indirect immunoenzymatic technique detecting TOXOPLASMA GONDII and its antigen in infected mice

目的:用间接免疫酶法检测小鼠组织内弓形虫及抗原成份,为实验研究和制备诊断试剂盒提供材料。方法:选择最佳的实验条件,包括羊抗兔IgG-HRP的浓度、反应时间的选择,观察了消化液对染色的影响,并对该方法的敏感性、特异性、重复性进行了研究。结果:羊抗兔IgG-HRP的最适浓度为1∶100。最适抗体抗原反应时间为24h,抗体与羊抗兔IgG-HRP反应时间为1h,最适显色时间为5min。间接酶法用于虫体涂片时,兔抗弓形虫血清1∶800为最高稀释度。在肝脏、脾、肺和脑均可清楚显示弓形虫感染组织内弓形虫速殖子及抗原成份,染为棕黄色。结论:本方法敏感性高、特异性强、重复性好。该方法可用于弓形虫病的病理诊断。

Objective： Using indirect immunoenzymatic technique to detect TOXOPLASMA GONDⅡ and its antigen in infected mice in order to provide material for experimental research and clinical diagnosis of toxop lasmosis. Methods： The best experimental conditions were selected, such as suitable response time, sheep an ti-rabbit HRP serum dilute concentration, effect of digest liquor to stain and have studied sensitivity, specific ity and repetition of the technique. Results： The suitable dilute concentration of sheep anti-rabbit HRP serum is 1：100. The proper time of the reactions between TOXOPLASMA GONDH antigen and antibody, antibody and sheep anti-rabbit HRP serum and staining is 24 h, 1 h, 5 min respectively. Using to stain toxoplasma smear preparation, the highest dilute concentration of rabbit anti- toxoplasma serum is 1：800. The dilute rabbit anti- toxoplasma serum can be kept 2 months under -20 ℃. In liver, spleen, lung and brain, the indirect immunoenzymatic technique can demonstrate tachyzoites, cyst and toxoplasma antigen clearly in the infected tissues. Conclusion： The technique has characteristics of high sensitivity,strong specificity and good repetition. The method can be used to make pathology diagnosis.