Determination of Serum and Urine Proteins by Electrophoresis and Its Importance in Clinical Veterinary Medicine


Abstract: In this study, for the investigation of routine clinical usage of urine protein electrophoresis in the laboratory to compare with the clinical findings in veterinary medicine, qualitative and quantitative changes in protein patterns of serum and urine from dogs with kidney disorders and proteinuria were investigated using electrophoresis. Eighteen dogs (patient group)of various ages and breeds with kidney disorders proved in the clinic and the laboratory and seventeen healthy dogs (control group)were used in this study. The histories (age,sex,breed)of the dogs in the control and patient groups were recorded in the clinic.The blood was collected from the cephalic vein,and the serum was removed using a centrifuge.Urine was obtained by a sterile catheter from the ureter.Biochemical analysis of urine was performed in the clinic.The urines of dogs with proteinuria were centrifuged and ultrafiltrated to concentrate for protein electrophoresis. Serum and urine proteins were separated by 10 %SDS-PAGE. Creatinine in serum and urine was detected using the Jaffe reaction, and urea and total protein in serum by the modified Gentzkow and biuret methods respectively. A/G ratios in serum and protein/creatinine ratios in urine were calculated arithmetically. Levels of serum urea and creatinine were found to be high and statistically significant (p<0.05 and p<0.01 respectively)in the diseased group. In the diseased group, 66 kDa serum protein pattern was significantly high (p<0.05); this may have a correlation with serum albumine in their size.In acute disorders,55 kDa serum protein pattern tended to increase. Disorders of three dogs with kidney disease were diagnosed by urine protein electrophoresis before the increased levels of urea and creatinine in serum.Urine proteins of eighteen dogs with primary kidney disease were separated by electrophoresis,and glomerular, glomerulo-tubular,tubular disorders were seen in six,four and four dogs respectively.These disorders were usually found to be acute when evaluated with the other laboratory and clinical findings. Protein patterns were divided into three groups by SDS-PAGE analysis;proteins with high molecular weight (>45 kDa)refer to glomeruler or glomerulo-tubular disorders,proteins with medium molecular weight (45-25 kDa)refer to tubulo-interstitial disorders, and proteins with small molecular weight (25-10 kDa)refer to tubular disorders. For the treatment of dogs with the disease,an antibiotic (sephalosporin,20 mg/kg and ampisilin,100 mg/kg)via intramuscular, and vit C (redoxan)and vit B complex (neurogriseovit)were applied.Ringer solution with lactate and 5 %dextrose were also applied intravenously to the dogs.There was no significant differences in the values of clinically significant various serum and urine parameters between pre-and post-treatments of dogs.However,the dogs with disease recovered well according to clinical and biochemical parameters after treatment. In conclusion,molecular weights and percentage distribution of urine protein patterns may be used routinely for the diagnosis of kidney diseases and to detect the location of disorders and although a small number of animals were used and analysed in a short time,it may also be useful for the prognosis and for the follow up of responses to the treatments.

Keywords: Dog, Kidney disorder, Proteinuria, SDS-PAGE, Urine, Serum.

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