首页> 中文期刊> 《中国人兽共患病学报》 >抗弓形虫速殖子单克隆抗体的制备与特性研究

抗弓形虫速殖子单克隆抗体的制备与特性研究

         

摘要

Aim To prepare and identify monodonal sntibody (Mab) specific for Toaoplasma gondii tachyzoites. Method The Mab specific for Taxoplasma gondii tachyrzoite were prepared via bybridoma technique. Indirect ELISA was used to determine the activity of the Mab. Agarose double immuodiffusion test was performed to confirm subclass and SDS-PAGE & western blot were used to analysis rolecular weight of the antigen (s) recognized by the Mab. IFA was used to identify the epitope of Taxoplasma gondii tachyzoites. The protection and specificity of the Mab were snalysed at same time. The Mab was tesed in Mab-ELISA method to detect Taxoplasma gondii antigen. Results A Msb F7C8H12 specific for T. gondii was produced. It belongs to IgG1 subclass. Moleculsr weight of the sntigens recognized by the Mab was 16.5 and 24 kDa. IFA did not show fiuorescence in intact tachyzoite.Inhibition test showed that the inhibition rate was 50% when the concetration of the antigen was 40μg/ml.Afterthe RH strain tachyzoites were incubated with Mab ascites, mice were inected with the tachyzoites through peritoneum. The results showed that the mean dead time of mice were not delayed. T. gondii antigen mixed with PBS snd normal human serum was detected by Mab-ELISA, the sensitvity was 0.78 yg/ml and 1.5μg/ml respectively. When mice were infected with T. gondiiRH strain tachyzoites, 103/mouse p.i., circulating antigen could be detectedin 6 day and 8 day. Conchusion The Mab (F7C8H12) to T. gomdii tachyzoites is an excellent probe for studying T. gondii snd toxoplasmosis.%目的 制备抗弓形虫速殖子单克隆抗体(Mab),并对其特性进行分析,探讨其在弓形虫病诊断中的作用。方法 利用杂交瘤技术制备抗弓形虫速殖子Mab。用间接ELISA法测定活性,以琼脂糖免疫双扩散鉴定亚类,通过SDS-PAGE和Westernblot分析该Mab所识别的抗原分子量,以IFA确定抗原位点在虫体的定位,并对其保护性及特异性进行分析。通过Mab-ELISA法检测弓形虫抗原,以研究其在弓形虫病血清学诊断中的意义。结果 抗弓形虫Mab F7C8H12属IgG1亚类,识别抗原的分子量为16.5和24kDa。以固定的完整虫体进行IFA,未显示荧光。竞争抑制试验显示50%抑制率时的抗原浓度为40μg/ml。将RH株速殖子与Mab腹水进行孵育后接种于小鼠腹腔,并不能延长小鼠的平均死亡时间。以Mab-ELISA法检测人工溶于PBS和正常人血清中的弓形虫RH株速殖子抗原,最小检出量为0.78μg/Ml和L5μg/ml。RH株速殖子108/只感染小鼠,分别于第6d和第8d能检出循环抗原。结论 抗弓形虫速殖子MabF7C8Hl2是研究弓形虫和弓形虫病的很好探针,其应用前景值得进一步研究。

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