A high-sensitivity immuno-chemiluminescence technique for detection of oligoclonal IgG and IgM in unconcentrated cerebrospinal fluid.
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.
Nelson
D
D
Department of Clinical Chemistry, Hospital of Helsingborg, Sweden.
Fredman
P
P
Börjeson
J
J
D004587:Electrophoresis, Agar Gel; D006801:Humans; D007124:Immunoenzyme Techniques; D007074:Immunoglobulin G / Q000134:cerebrospinal fluid*; D007075:Immunoglobulin M / Q000134:cerebrospinal fluid*; D008163:Luminescent Measurements; D012016:Reference Values
D007074:Immunoglobulin G; D007075:Immunoglobulin M