中国寄生虫学与寄生虫病杂志 ›› 2001, Vol. 19 ›› Issue (6): 12-362.

• 论著 • 上一篇    下一篇

从感染小鼠获得微小隐孢子虫卵囊方法的改进

黄克和 1;杨世广2;唐建霞 1
  

  1. 南京农业大学动物医学院;Department of Animal;南京农业大学动物医学院 南京210095;Dairyand Veterinary Sciences;UtahState University;UT84322;USA;南京210095
  • 收稿日期:1900-01-01 修回日期:1900-01-01 出版日期:2001-12-30 发布日期:2001-12-30

Improvement of Amplification Method for Cryptosporidium parvum Oocysts from Mice

HUANG Ke he 1;YANG Shi guang 2;TANG Jian xia 1   

  1. 1 College of Veterinary Medicine;Nanjing Agricultural University;Nanjing 210095; 2 Department of Animal;Dairy and Veterinary Sciences;Utah State University;UT 84322;USA
  • Received:1900-01-01 Revised:1900-01-01 Online:2001-12-30 Published:2001-12-30

摘要:   目的 建立从感染C5 7BL/ 6N小鼠获得大量微小隐孢子虫卵囊的方法。方法 在饮水中添加不同浓度的地塞米松对人工感染小鼠进行免疫抑制 ,探讨获得最多卵囊数的药物添加量 ;比较不同纯化方法得到的卵囊回收率和纯度 ;探讨来自不同动物、不同纯化方法的卵囊在牛输卵管上皮细胞培养系统的感染情况。结果 地塞米松加入量为 2 0 μg/ml的第 3组获得的卵囊量高达 4 .16× 10 9/ 30个小鼠 ;用饱和盐水漂浮法与用蔗糖密度梯度离心法纯化得到的卵囊具有相同的回收率和纯度 ;从感染小鼠与犊牛中获得的卵囊在感染性上差异不显著。结论 成功地建立了从感染小鼠获得大量微小隐孢子虫卵囊的方法 ,具有简单、方便、快速等优点 ,并且不影响活力。

关键词: 微小隐孢子虫卵囊, 小鼠, 饱和盐水漂浮法, 蔗糖密度梯度离心法, 细胞培养

Abstract:  Objective To establish a simplified amplification method for obtaining a large number of purified Cryptosporidium parvum oocysts from infected C57BL/6N mice. Methods All mice in the experimental groups were immunosuppressed by given different concentrations of dexamethasone phosphate added in drinking water throughout the experiment. The recovery and purity of the oocysts obtained using different purification methods was compared. The infectivity of the oocysts obtained from the same origin but different animals and different purification methods in a bovine fallopian tube epithelial cell culture system was studied. Results 4.16×10 9 oocysts were obtained in 30 mice in the 3rd group with dexamethasone of 20 μg/ml in drinking water. No significant difference in the oocyst recovery, purity and infectivity was found between methods using saturated saline floatation and sucrose density gradient centrifugation. The infectivity of the oocysts obtained from the same origin but different animals was similar. Conclusion A simplified amplification method for obtaining a large number of purified Cryptosporidium parvum oocysts from the infected mice was established.

Key words: Cryptosporidium parvum oocysts, mouse, saturated saline floatation, sucrose density gradient centrifugation, cell culture