中国寄生虫学与寄生虫病杂志 ›› 1984, Vol. 2 ›› Issue (2): 88-90.

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弓形虫病的血清学诊断研究 Ⅰ.弓形虫速殖子的纯化

杨惠珍,徐克继   

  1. 上海第二医学院寄生虫学教研室
  • 收稿日期:2017-01-12 修回日期:2017-01-12 出版日期:1984-05-31 发布日期:2017-01-12

STUDIES ON THE SEROLOGICAL DIAGNOSIS OF TOXOPLASMOSIS I. PURIFICATION OF TOXOPLASMA GONDII TACHYZOITES

  • Received:2017-01-12 Revised:2017-01-12 Online:1984-05-31 Published:2017-01-12

摘要: 本文参照Tsunematsu(1960)及Petrov(1974)方法,用弓形虫猪株和人株感染小鼠收集腹腔液,经胰蛋白酶消化,G_3砂芯漏斗过滤,离心,制备较纯的虫体抗原,并用冻干法长期保存。对数批感染小鼠的腹腔液在处理前、后分别进行虫体和细胞计数,用伊文思蓝液染色鉴别死活。并测定虫体的得量、纯度,存活率和回收率。弓形虫猪株和人株感染小鼠腹腔液经上述处理后平均每只获虫体分别为(7.9±1.3)×10~6和(7.5±1.7)×10~6,两者得量无显著差异(t=0.55,P0.05);虫体纯度在处理前为0.688±0.18, 处理后为0.993±0.004有显著性差异(t=8.125,P0.01)

关键词: 弓形虫病, 血清学, 速殖子, 腹腔液, 诊断研究, 存活率, 回收率, 感染小鼠, 酶消化, 伊文思蓝

Abstract: A modification of the procedure described by Tsunematsu (1960) and Petrov (1974) for purification of Toxoplasma parasites ,was reported. Both pig and human strains of T. gondii maintained in mice by peritoneal inoculation were used. For parasite purification, peritoneal fluids were collected, digested with trypsin and filtered through a glass filter (G3) to remove host cells. Suspended parasites were freeze-dried for longterm preservation. Parasites and host cells were counted before and after digestion in some batches of infected mice, and Evan's blue staining was adopted for the examination of parasite vitality. Parasite yield, purity, viability as well as recovery rate were determined and compared in different lots.Application of this modified procedure resulted in an average parasite yield of (7.9±1.3) ×106 and (7.5±1.7) × 106 per mouse in pig and human strains respectively with no statistical difference (t = 0.55; p0.05). A high purity of 0.993±0.004 after treatment versus 0.668±0.18 before treatment was obtained, the noted difference being highly significant(t= 8.125; p0.01). The mean recovery rate as calculated from 4 lots has been 87.5% (t = 1.33; p0.05).