卡氏肺孢子虫包囊抗原纯化方法的研究

STUDY ON PURIFICATION OF CYST ANTIGEN OF PNEUMOCYSTIS CARINII

目的 研究卡氏肺孢子虫包囊抗原的纯化方法。 方法 肌肉注射地塞米松诱发大鼠卡氏肺孢子虫肺炎 ,瑞 -姬氏染液复合染色法检获肺组织中的肺孢子虫包囊 ,用改进的胶原酶消化法、Percoll不连续密度梯度离心分离、超声粉碎纯化卡氏肺孢子虫包囊抗原。 结果 采用浓度为 0 .0 5 %胶原酶消化 2次 ,30 min/次 ,在 1.0 30 g/ ml密度层可获得相对纯净而完整的卡氏肺孢子虫包囊抗原。 结论 本研究可提供高纯度卡氏肺孢子虫抗原 ,为进一步建立对卡氏肺孢子虫肺炎的有效诊断方法具有十分重要的意义。

Objective To study purification of cyst antigen (CAg) of Pneumocystis carinii . Methods Rats afflicted with P. carinii pneumonia were injected intramuscularly with dexamethasone. The cysts of P. carinii were examined by Wright Giemsa's Compound Stain. The pure CAg was obtained using improved digestion with collagenase, separation via a discontinuous density gradient centrifugation in Percoll and ultrasonication. Results The fragments of lungs were digested twice with 0.05% collagenase for 30 min, after which the intact and pure cysts were concentrated at the band of 1.030 g/ml. Conclusion The pure CAg obtained by the above process may be an useful technique in creating an effective future diagnostic method for P. carinii pneumonia.