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[利用纤维素膜过滤法纯化刚地弓形虫速殖子及制备可溶性抗原]

[Purification of Toxoplasma gondii Tachyzoites with cellulose membrane filtration and preparation of soluble antigen].

作者信息

Fang Zheng-Ming, Wang Sheng, Su Bin-Tao, Gong Nian-Qiao, Liu Wen-Qi, Ming Chang-Sheng

机构信息

Department of Parasitology, Tongji Medical College, Huazhong University of Science and Technology, Wahan 430030, China.

出版信息

Zhongguo Ji Sheng Chong Xue Yu Ji Sheng Chong Bing Za Zhi. 2010 Oct 30;28(5):329-31.

Abstract

20 ml peritoneal lavage fluid of mice infected with Toxoplasma gondii RH strain was diluted to 250 ml with sterilized physiological saline, and filtered through cellulose membrane filters (pore size: 5 microm). The filtrate was centrifuged at 1512 x g for 15 min, and the sediment was pure T. gondii tachyzoites which were then sonicated. The soluble antigen was prepared by centrifugation at 11200 x g for 30 min. Sera of T. gondii infected SD rat and normal SD rats were collected for immunodetection of soluble antigen. The specificity and valence of soluble antigen were detected with indirect ELISA. The mean removal rates of mouse leukocytes and erythrocytes were 99.9% and 80.3%, respectively, and recovery rate of tachyzoites was 71%. The soluble antigen was extracted from purified T. gondii (1.38 mg per mouse). Indirect ELISA showed that the lowest effective antigen concentration was 5 microg/ml.

摘要

将感染弓形虫RH株的小鼠20 ml腹腔灌洗液用灭菌生理盐水稀释至250 ml,经纤维素膜过滤器(孔径:5微米)过滤。滤液在1512×g下离心15分钟,沉淀物为纯弓形虫速殖子,然后进行超声处理。通过在11200×g下离心30分钟制备可溶性抗原。收集感染弓形虫的SD大鼠和正常SD大鼠的血清用于可溶性抗原的免疫检测。用间接ELISA检测可溶性抗原的特异性和效价。小鼠白细胞和红细胞的平均去除率分别为99.9%和80.3%,速殖子回收率为71%。从纯化的弓形虫中提取可溶性抗原(每只小鼠1.38 mg)。间接ELISA显示最低有效抗原浓度为5μg/ml。

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