Improved detection of oligoclonal IgG in cerebrospinal fluid by isoelectric focusing in agarose, double-antibody peroxidase labeling, and avidin-biotin amplification.
To demonstrate oligoclonal IgG bands (I) in unconcentrated cerebrospinal fluid, we used isoelectric focusing in agarose followed by protein transfer to cellulose nitrate membrane, double-antibody peroxidase labeling, and avidin-biotin amplification. I can be reliably seen after isoelectric focusing of 5-microL specimens containing 125 ng of IgG (25 mg/L). Thus the technique is more sensitive than others (e.g., silver staining) and more reliable than radioimmunofixation. When we used this technique with fluids from 62 patients with multiple sclerosis and infectious disease of the central nervous system, 84% displayed I, a percentage not increased when the same specimens were concentrated to 3.5 g of IgG per liter, examined by agarose isoelectric focusing, and stained with Coomassie Blue. Results for 53 patients with tension headache and psychoneurosis were all negative. By obviating the need to concentrate samples of cerebrospinal fluid the present method is a useful, sensitive alternative for demonstrating I.
D001360:Avidin; D001710:Biotin; D002493:Central Nervous System Diseases / Q000276:immunology; D003101:Collodion; D005455:Fluorescent Antibody Technique; D006801:Humans; D007124:Immunoenzyme Techniques; D007074:Immunoglobulin G / Q000134:cerebrospinal fluid* / Q000302:isolation & purification; D007202:Indicators and Reagents; D007239:Infections / Q000276:immunology; D007525:Isoelectric Focusing / Q000379:methods; D009103:Multiple Sclerosis / Q000276:immunology; D012394:Rosaniline Dyes; D012685:Sepharose
D007074:Immunoglobulin G; D007202:Indicators and Reagents; D012394:Rosaniline Dyes; D001360:Avidin; D001710:Biotin; D003101:Collodion; D012685:Sepharose; C004692:coomassie Brilliant Blue