›› 2010, Vol. 28 ›› Issue (5): 15-331.

• 研究简报 • Previous Articles     Next Articles

Construction and identification of a full-length cDNA library from Spirometra erinaceieuropaei

FANG Zheng-ming1，WANG Sheng2，SU Bin-tao1，GONG Nian-qiao2，
LIU Wen-qi1，MING Chang-sheng2 *   

  1. 1 Department of Parasitology，Tongji Medical College，Huazhong University of Science and Technology，Wuhan 430030，China；2 Institute of Organ Transplantation，Tongji Hospital，Tongji Medical College，Huazhong University of Science and Technology，Wuhan 430003，China
  • Received:1900-01-01 Revised:1900-01-01 Online:2010-10-30 Published:2010-11-09
  • Contact: MING Chang-sheng

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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 μm）. The filtrate was centrifuged at 1 512×g for 15 min， and the sediment was pure T. gondii tachyzoites which were then sonicated. The soluble antigen was prepared by centrifugation at 11 200×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 μg/ml.

Key words: Toxoplasma gondii, Purification, Antigen, Cellulose membrane filter