安氏隐孢子虫卵囊分离纯化方法研究

目的 建立获取纯净的安氏隐孢子虫卵囊外培养模型。方法研究改进分离纯化安氏隐孢子虫卵囊的方法，采用蒸馏水稀释Sheather’s蔗糖溶液配制不同密度的蔗糖溶液，比较不同稀释比的Sheather’s蔗糖溶液漂浮卵囊的效果，并首次比较了蒸馏水和PBS稀释的Sheather’s蔗糖溶液分离纯化卵囊的活性，将两种方法分离纯化的卵囊37℃水浴1h进行脱囊率比较。结果2：1蔗糖溶液（2体积Sheather’s蔗糖溶液与1体积的蒸馏水）分离奶牛粪样中卵囊效果最好。漂浮三次即可获得大量卵囊，回收率为83．70％，三次漂浮的卵囊总数较Sheather’s蔗糖溶液高57．53％。蔗糖溶液多梯度密度纯化法纯化后的卵囊杂质较少，回收率为47．52％。蒸馏水稀释的蔗糖溶液分离纯化卵囊脱囊率为87．87％，PBS稀释的蔗糖溶液分离纯化卵囊的脱囊率为84．71％，差异无统计学意义P〉0.05）。结论2：1蔗糖溶液适用于分离安氏隐孢子虫卵囊，且用蒸馏水代替PBS稀释Sheather’s蔗糖溶分离纯化隐孢子虫卵囊其活性不受影响。可用于体外培养研究。

Methodological study on the isolation and purification of Cryptosporidium andersoni oocysts

Objective To obtain Cryptosporidium andersoni oocysts from feces to culture in vitro.Methods The present study was undertaken to isolate and purify Cryptosporidium andersoni oocysts using different diluted Sheather's solution with distilled water and PBS, in which recovery rates and excystation rates were compared.Results The recovery rate was 83.70% when oocysts were isoalted by 2:1Sherther's sucrose solution diluted with distilled water in three times, and the number of oocysts was 57.53% more than the nu...