Popov G, Martinov S, Dimitrova Z
Vet Med Nauki. 1981;18(6):3-11.
A method was worked out for the production of an antichlamydial fluorescent serum to be used in the direct immunofluorescence test. Used were the methods of culturing Chlamydia in chick embryos and cell cultures, light and electron microscopy, differential centrifugation and supercentrifugation, gel chromatography in Sepharose, ultracentrifugation in saccharose, hyperimmunization of guinea pigs, conjugation of gammaglobulin with fluorescein isothiocyanate, complement fixation, and the direct immunofluorescence itself. Applied was a new immunologic scheme for the production of fluorescent sera. The guinea pigs were hyperimmunized twice and three times at 30-day intervals with highly purified, highly concentrated and adjuvant-treated chlamydial suspensions. It was concluded that the main condition for the production of high titer labelled gammaglobulins was to obtain purified and concentrated chlamydial antigens and use these with an adjuvant. To indicate Chlamydia in testing the conjugates it is essential to prepare standard test lamellas infected at high multiplicity of purified and concentrated Chlamydia.
制定了一种生产用于直接免疫荧光试验的抗衣原体荧光血清的方法。所采用的方法包括在鸡胚和细胞培养物中培养衣原体、光学和电子显微镜检查、差速离心和超速离心、琼脂糖凝胶色谱法、蔗糖超速离心、豚鼠超免疫、γ球蛋白与异硫氰酸荧光素偶联、补体结合以及直接免疫荧光本身。应用了一种生产荧光血清的新免疫方案。以高度纯化、高度浓缩并经佐剂处理的衣原体悬液，每隔30天对豚鼠进行两次和三次超免疫。得出的结论是，生产高滴度标记γ球蛋白的主要条件是获得纯化和浓缩的衣原体抗原，并将其与佐剂一起使用。为了在检测偶联物时指示衣原体，制备在高感染复数下感染纯化和浓缩衣原体的标准测试片至关重要。
