Manych J
J Hyg Epidemiol Microbiol Immunol. 1984;28(2):207-16.
Up to now, the diagnosis of candidiases has been an intriguing problem chiefly because the detection of Candida in the sick organism is no conclusive evidence of its etiological involvement. The endeavour to use serological methods in the diagnosis has not been successful and their use has so far practically been limited to taxonomic studies. In an effort to find a serological approach to the diagnosis, we found the agarimmunodiffusion assay of Ouchterlony to be the most promising method and used it in our own modification. The first stage of the work was to select and prepare the most suitable type of antigen for this reaction (described in the present paper). Sixteen antigen types classifiable into four groups were prepared: simple water-extracted antigens, alcohol-extracted antigens, antigens obtained on disrupting cells by repeated freezing and thawing, and antigens prepared by boiling cultures. Cultures for antigen preparation were grown on Salvin's medium or on liquid Sabourad's medium and the antigens obtained were either used in their native form or dialysed. Serological tests with hyperimmune rabbit sera prepared by our own schedule were done repeatedly. By far the best were simple water-extracted antigens, nondialysed, from cultures grown on Sabouraud's medium. They reacted the most sensitively, gave high antibody detection rates and assessable precipitin reactions and showed high species specificity. Tests with positive human sera fully confirmed these findings; in fact, species specificity was even somewhat higher here.
到目前为止,念珠菌病的诊断一直是个棘手的问题,主要原因是在患病机体中检测到念珠菌并不能确凿证明其与病因有关。尝试使用血清学方法进行诊断并未成功,目前其应用实际上仅限于分类学研究。为了找到一种血清学诊断方法,我们发现Ouchterlony琼脂免疫扩散试验是最有前景的方法,并对其进行了我们自己的改进。工作的第一阶段是为该反应选择并制备最合适的抗原类型(本文对此进行了描述)。制备了可分为四组的16种抗原类型:简单水提取抗原、醇提取抗原、通过反复冻融破坏细胞获得的抗原以及煮沸培养物制备的抗原。用于制备抗原的培养物在Salvin培养基或液体Sabouraud培养基上生长,获得的抗原要么以天然形式使用,要么经过透析。用我们自己制定方案制备的超免疫兔血清反复进行血清学检测。到目前为止,最好的是来自在Sabouraud培养基上生长的培养物的未透析的简单水提取抗原。它们反应最灵敏,抗体检测率高,沉淀素反应可评估,并且具有高种属特异性。用人阳性血清进行的检测充分证实了这些发现;事实上,这里的种属特异性甚至更高一些。