Nelson D, Fredman P, Börjeson J
Department of Clinical Chemistry, Hospital of Helsingborg, Sweden.
Scand J Clin Lab Invest. 1994 Feb;54(1):51-4. doi: 10.3109/00365519409086509.
A method is described for detection of oligoclonal IgG and IgM in unconcentrated cerebrospinal fluid (CSF). The proteins were separated by agarose gel electrophoresis and transferred to cellulose nitrate membranes. A double antibody technique with a peroxidase conjugated secondary antibody was used to detect the protein. Bound secondary antibody was visualized by autoradiography using chemiluminescence (ECL) reagent. The procedure presented here allows examination of oligoclonal bands of IgG and IgM in less than 6h. The detection limit for a single band was approximately 10pg and the results were in complete agreement with those achieved by isoelectric focusing (Phast System) and subsequent immunofixation. In 9 of the samples oligoclonal IgM was also detected.
本文描述了一种检测未浓缩脑脊液(CSF)中寡克隆IgG和IgM的方法。蛋白质通过琼脂糖凝胶电泳分离,然后转移至硝酸纤维素膜上。采用带有过氧化物酶标记二抗的双抗体技术检测蛋白质。使用化学发光(ECL)试剂通过放射自显影使结合的二抗可视化。此处介绍的方法能够在不到6小时内检测IgG和IgM的寡克隆条带。单一条带的检测限约为10 pg,结果与等电聚焦(Phast系统)及随后的免疫固定法完全一致。在9个样本中还检测到了寡克隆IgM。