Lee K H, Shin W S, Park H S, Park J Y, Koh C M
Department of Microbiology, Yonsei University Wonju College of Medicine, Kangwon-Do, Korea.
Yonsei Med J. 1997 Jun;38(3):178-86. doi: 10.3349/ymj.1997.38.3.178.
From the culture filtrates of C. albicans, C. tropicalis and C. parapsilosis, proteinases were purified using a series of chromatographic steps consisting of DEAE-Sepharose, Sephacryl S-200 and size-exclusion HPLC which removed contaminating mannoproteins and extraneous proteins. Anti-Candida proteinase antibodies in sera from mice infected with various Candida species were detected using ELISA for serodiagnosis of candidiasis. Three proteinases were blotted by homologous and heterologous anti-proteinase antisera on Western blot analysis. All sera from six Candida species-infected mice were reactive with proteinases of C. albicans, C. tropicalis, and C. parapsilosis, although C. glabrata, C. guilliermondii, and C. krusei did not secrete proteinase. The seroreactivities of proteinase with sera from mice infected with homologous C. albicans and C. tropicalis were higher than those with sera from heterologous Candida species-infected mice. These results suggest that three proteinases have at least one common epitope, but its application for diagnosis of candidiasis should be considered with limits of specificity.
从白色念珠菌、热带念珠菌和近平滑念珠菌的培养滤液中，通过一系列色谱步骤（包括DEAE-琼脂糖、Sephacryl S-200和尺寸排阻HPLC）纯化蛋白酶，这些步骤可去除污染的甘露糖蛋白和外来蛋白。使用酶联免疫吸附测定（ELISA）检测感染各种念珠菌属的小鼠血清中的抗念珠菌蛋白酶抗体，用于念珠菌病的血清学诊断。在蛋白质印迹分析中，三种蛋白酶被同源和异源抗蛋白酶抗血清印迹。来自感染六种念珠菌属小鼠的所有血清均与白色念珠菌、热带念珠菌和近平滑念珠菌的蛋白酶发生反应，尽管光滑念珠菌、季也蒙念珠菌和克柔念珠菌不分泌蛋白酶。蛋白酶与感染同源白色念珠菌和热带念珠菌的小鼠血清的血清反应性高于与感染异源念珠菌属小鼠血清的反应性。这些结果表明，三种蛋白酶至少有一个共同表位，但其在念珠菌病诊断中的应用应考虑特异性的局限性。
