The Research Of The Diagnostic Significance Of Kidney Damage By Using Immunofixation Electrophoresis Method To Detect The Urine Protein

Urine protein is a chemical component of urine, it means one excrete more than 0.20g/L protein each day. In normal, one may excrete about 50-150mg protein to urine each day, most of the protein is albumin. Various pathopoiesis factor can destroy the unit of the kidney, damage the glomerular filtration membrane barrier and/or electric charge barrier, the plasma protein in urine increased and the reabsorption of it decreased, then the urine protein come about. The urine protein is a common clinical manifestation of the kidney damage, so the detection of the urine protein become a important index for diagnosis,differential diagnosis and curative effect of kidney disease. It is about 300 years from 1695 when Dekkers use acidification and heating method to confirm that the urine contains proteins. During these years many specialist both in domestic and in abroad got many effective methods to detect the protein in urine, such as alkaloid urine protein quantitative method, which is used as a routine in clinic, but these methods have many shortage, such as can not direct-viewing and high cost. In order to construct a direct-viewing,clear,fast and low cost method, our research according to the principle of immunity and electrophoretic and using agarose gel as the sustentaculum to construct a high tension and tachyphylaxis fixiation electrophoresis method to detect the urine protein.Principle of the experiment: In combination of electrophoretictechnology and immunological reaction between antigen and antibody, the diffusion of antigen is accelerated by the electric field, and its existence is confirmed by specific antibody. Based on the agarose gel as the carrier, proteins with different molecular weight and isoelectric point are separated according to the different electrophoretic velocity. Then monoclonal antibodies with different molecular weight were added to the gel, and the identification of the protein is achieved with the specificity of antigen-antibody reaction.Procedure of the experiment: The urine samples were run on the agarose gel at the circumstance of high voltage and hypothermia, with four zones each sample. Proteins in the first zone were precipitated with stationary liquid, and GAM, MACRO, MICRO monoclonal antibodies were applied to sediment the proteins in the other three zones respectively. Sensitive dye was used to stain the gel. The proteinuric type and degree of kidney damage were defined by analysis of zones on the gels.Four protein zones were harvested by immunofixation electrophoresis of 120 urine protein positive samples and 30 samples from healthy controls, corresponding to physiological proteinuria, glomerular proteinuria, tubuloproteinuria and mixed proteinuria. Glomerular proteinuria was classified into selective proteinuria, intersexus proteinuria, nonselective proteinuria. It is helpful in locating the site and defining the extent of kindy damage.High voltage rapid immunofixation eletrophoresis ischaracterized by high sensitivity, high specificity, time-saving, result of diret-viewing and analysis easily performed. With these advantages, it actually is an good method for diagnosis and differential diagnosis of proteinuria with potential value of clinical application.