国产泛影葡胺密度梯度沉降法纯化Q热立克次体

Q热立克次体Henzerling株鸡胚培养的感染卵黄囊在蔗糖-磷酸缓冲生理盐水中制成悬液,以差速离心法提取,继用复方泛影葡胺密度梯度离心法排除鸡胚卵黄囊成分,提取和纯化活的立克次体。在线性密度梯度液中段,立克次体形成乳白色界限明晰的区带。 经光学显微镜及电子显微镜检查,立克次体的形态完整。在豚鼠、小白鼠及鸡胚感染实验中表明仍保留着活力及感染性。免疫荧光试验,单克隆抗体保护试验及补体结合试验证明对Ⅰ相及Ⅱ相的抗原性无变化。此提纯的制剂中未查出污染的鸡胚卵黄囊成分,其平均回收量为0.868mg/g卵黄囊或0.3472mg/g卵黄囊。

The purification of coxiella burnetii by meglumini diatrizoatis composita density gradient centrifugation

Coxiella burnetii ( Henzerling strain), cells cultivated in the yolk sac of chicken embryo.es were separated from host cell components by the Meglumini diatrizoatis density gradient centrifugation. Initial step in the purification of viable rickettsia was differential centrifugation. Then it was separated and formeda clear-cut band by the gradient centrifugation.Among the suspension of the purified viable Coxiella burnetii the inte grity of morphologically different cell variants were maintained during the purification procedure.The activity and infectivity for guinea pigs,mice and chicken embryoes were retained. It was proved that antigenicity of Coxiella burnetii phase I and II were still actively reacted in immunofluorescent technic, monoclonal antibody technic and Complement fixation test.It was free from host cell components and the average yield of Coxiella burnetii was 0.868 mg per each yolk sac or 0.3472 mg per gram of yolk sac.