Poor A H, Cutler J E
J Clin Microbiol. 1979 Mar;9(3):362-8. doi: 10.1128/jcm.9.3.362-368.1979.
The development of a solid-phase radioimmunoassay procedure for the detection of Candida albicans antigens in serum of mice is described. Antibodies against C. albicans that were used in the radioimmunoassay procedure were partially purified from immune serum by a C. albicans antigen-coupled affinity column. Elution of anti-C. albicans antibodies from the column was by glucose and mannose; 4 mg of protein was recovered per ml, which contained 50% of the candidal agglutinin activity of immune serum. Also, 81% of the protein (partially purified antibody) recovered was adsorbed by whole C. albicans cells. Anti-C. albicans antibodies were either coupled to Sepharose 4B for use as the solid phase to bind candidal antigen in serum of infected animals, or radioiodinated (125I) for use as a tracer molecule to bind to the candidal antigen solid-phase complex. Although control experiments indicated that at least 100 ng of candidal antigen should be present in a serum specimen for a positive radioimmunoassay test, candidal antigenemia was detected in 70.4% of infected mice even in cases where blood cultures for C. albicans were negative. With further refinement and adaptability to human serum, the radioimmunoassay test may become a helpful tool for use in the diagnosis of systemic candidiasis.
本文描述了一种用于检测小鼠血清中白色念珠菌抗原的固相放射免疫分析方法的开发。用于放射免疫分析的抗白色念珠菌抗体通过白色念珠菌抗原偶联亲和柱从免疫血清中部分纯化。抗白色念珠菌抗体从柱上的洗脱采用葡萄糖和甘露糖;每毫升回收4毫克蛋白质,其含有免疫血清50%的念珠菌凝集素活性。此外,回收的蛋白质(部分纯化抗体)的81%被完整的白色念珠菌细胞吸附。抗白色念珠菌抗体要么偶联到琼脂糖4B上用作固相,以结合感染动物血清中的念珠菌抗原,要么进行放射性碘化(125I)用作示踪分子,以结合念珠菌抗原固相复合物。尽管对照实验表明,血清标本中至少应存在100纳克念珠菌抗原才能进行阳性放射免疫分析检测,但即使在白色念珠菌血培养为阴性的情况下,仍在70.4%的感染小鼠中检测到念珠菌抗原血症。随着进一步改进并适用于人血清,放射免疫分析检测可能成为诊断系统性念珠菌病的有用工具。