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A new screening method for proteinuria using Erythrosin B and an automated analyzer--rapid, sensitive and inexpensive
Satoshi Horikoshi1, Asami Higurashi, Emiko Kaneko
1Division of Nephrology, Department of Internal Medicine, Juntendo University Faculty of Medicine, Tokyo, Japan.
Background:
In spite of the urgent necessity for a screening test of urinary protein for the early diagnosis of kidney diseases, a rapid, accurate and cost-effective method for their detection has yet to be developed.
Methods:
A solution containing a buffer agent (pH 2.3) and surfactants and a solution of Erythrosin B are added to a urine sample. After letting the mixture stand for 5 min at 37°C, the dye-bound protein is measured by a spectrophotometer at 546 nm using a Hitachi 7170S automated analyzer.
Results:
The calibration curve was linear with human serum albumin concentration in the range of 2.4-200 mg/l. The detection limit, 2.4 mg/l was superior to conventional dye-binding methods by one order of magnitude and comparable to the turbidimetric immunoassay (TIA). Spot urine samples from 70 patients who showed (-) or (±) in the dip-stick screening test for proteinuria and 79 healthy volunteers were analyzed. There was an excellent correlation (r=0.978, n=149) between the results given by the proposed method and those by the TIA.
Conclusions:
This method provides a viable alternative to the conventional immunoassay-based methods for urinary protein measurement, and will be useful in the diagnosis of early stage kidney disease.
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Enzyme-Linked Immunosorbent Assay
There are many different types of ELISAs, but they all involve an antibody molecule whose constant region binds an enzyme, leaving the variable region free to bind its specific antigen. Enzyme-substrate reaction allows the antigen to be visualized or quantified.

