摘要
20只纯系Wistar大鼠经皮下注射醋酸可的松25m/次,每wk2次,用药7wk后即可诱发大鼠肺孢子虫肺炎。经相差显微镜直接检查、姬姆萨染色、亚甲胺蓝染色及六亚甲基四胺银染色在肺组织内检出肺孢子虫包囊。分离纯化过程包括三个步骤:1 肺组织匀浆的制备及过滤;2 采用0.2％胶原酶消化;3 不连续Percoll密度梯度离心。包囊及滋养体密度为1.033g/ml,肺组织碎片密度达1.040g/ml,从而分离。经上述过程处理后可获得较为纯净的肺孢子虫包囊,可用于ELISA。
Twenty Wistar rats were injected with cortisone acetate twice a week subcutaneously for 6-12 wks.By 7 weeks Pneumocystis carinii pneumonia was induced in rats successfully.The cysts of Pneumocystis carinii from infected lungs of cortisonized rats were indentified by phase-contrast microscopy,Giemsa's stain,Chalvardjian's stain and Gomori's methenamine silver nitrate stain.The process of isolation and purification of Pneumocystis carinii from infected rat lungs included the following three test steps.First,the tissue was cut into small pieces and homogenized and filtered through #60,#100,#200-gauge wire mesh respectively.Secondly,the homogenate was digested with collagenase,the optimal concentration of collag-enase being 0.2%.Thirdly,the discontinuous percoll density gradient centrifugation was used to separate P.carinii cysts.The majority of P carinii cysts were present in a density zone of approximately 1.033g/ml and were essentially free from host lung debris,the latter being removed due to their higher density,1.040g/ml.
出处
《中国寄生虫学与寄生虫病杂志》
CAS
CSCD
北大核心
1992年第3期166-170,共5页
Chinese Journal of Parasitology and Parasitic Diseases
基金
重庆市科委博士小额资助项目
关键词
肺孢子虫病
分离
纯化
肺孢子虫
Pneumocystis carinii,Gomori's methenamine silver nitrate stain,isolation and purification,density gradient centrifugation.
