Deutsch 
Test-Category
ELISA
Pack size
96 Tests
Possible sample matrix and volumes
| Urine | 100 µl |
Incubation periods
Indications
Regulatory Status
Der ReFiNE® Dkk3 ELISA is an enzyme-linked immunosorbent assay (ELISA) intended for the quantitative determination of Dickkopf-related protein 3 (DKK3) in human urine samples. The assay is an in vitro diagnostic medical device and intended to be used by professional users in a laboratory environment. It can be performed manually or using an automated platform. The assay functions as an aid in the diagnosis of acute and/or chronically progressive renal impairment.
ReFiNE® Dkk3 – “TROPONIN“ OF THE KIDNEY AND NEW MARKER OF CHRONIC KIDNEY DISEASE PROGRESSION
Dickkopf-3 (DKK3), a member of a family of Wnt-pathway modulators [1], has been identified as a stress-induced, tubular epithelial cell-derived regulator of kidney fibrosis [2]. Genetic deletion, antibody-mediated blockade and tubular epithelial cell-specific abrogation of DKK3 reduced tubulointerstitial fibrosis in various mouse models of chronic kidney disease (CKD) and hence preserved kidney function [2]. Mechanistically, Dkk3 deficiency caused diminished canonical Wnt/ß-catenin signaling in tubular epithelial cells under pathological conditions, accompanied by amplified anti-fibrogenic inflammatory response within the injured kidney. DKK3 protein was detected in the urine of patients suffering from different types of CKD, correlating specifically and quantitatively with the magnitude of tubulointerstitial fibrosis in kidney biopsy specimens [2,3]. Tubulointerstitial fibrosis is the common pathological manifestation of etiologically different CKD that finally result in kidney failure. Thus, DKK3 in urine can be used as a diagnostic marker of progressive CKD. This was confirmed in a large patient cohort with various causes of CKD, where urinary DKK3 levels were independently associated with a significant decline in kidney function (glomerular filtration rate, GFR) over the next 12 months, regardless of the cause of kidney injury [3]. Moreover, studies in subjects with clinically inapparent kidney damage revealed that even moderately increased levels of DKK3 in the urine – as a sign of ongoing tubular cell stress – signify increased risk for future acute kidney failure and progressive loss of kidney function thereafter [4]. The present enzyme-linked immune sorbent assay (ELISA) is intended for the quantitative determination of DKK3 in human urine. In this respect it serves as a specific measuring tool for “tubular stress” and thereby for the quantification of the risk of future kidney function loss resp. CKD progression [3-8].
Test Principle
The ReFiNE® Dkk3 ELISA is a sandwich ELISA for the quantitative measurement of DKK3 in human urine. The wells of the microtiter plate are coated with a monoclonal anti-human-DKK3 capture antibody. During the first incubation step, DKK3 present in the sample is bound to the immobilized capture antibody, forming the first antigen-antibody complex. After a washing step to remove all unbound sample components, a horseradish peroxidase (HRP)-labeled detection antibody is added. A capture antibody-antigen detection antibody complex is formed. Unbound substances are removed by another washing step. After the addition of an enzymatic reaction substrate (tetramethylbenzidine, TMB), HRP catalyzes the oxidation of the colorless TMB into a blue, oxidized TMB. The reaction is stopped by adding an acidic stop solution, and the color changes from blue to yellow. The intensity of the yellow color is directly proportional to the concentration of DKK3. Results are determined via a calibration curve.
References:
[1] Glinka, A., et al.: Dickkopf-1 is a member of a new family of secreted proteins and functions in head induction. Nature 1998; 391: 357-362.
[2] Federico, G., et al.: Tubular Dickkopf-3 promotes the development of renal atrophy and fibrosis. JCI Insight 2016; 1: e84916.
[3] Zewinger, S., et al.: Dickkopf-3 (Dkk3) in urine identifies patients with short-term risk of eGFR loss. J Am Soc Nephrol 2018; 29: 2722-2733.
[4] Schunk, S.J., et al.: Association between urinary dickkopf-3, acute kidney injury, and subsequent loss of kidney function in patients undergoing cardiac surgery: an observational cohort study. Lancet 2019; 394: 488-496.
[5] Sanchez-Alamo, B., et al.: Urinary Dickkopf-3: a new biomarker for CKD progression and mortality. Nephrol Dial Transplant 2021; 36: 2199-2207.
[6] Schunk, S.J., et al.: Measurement of urinary Dickkopf-3 uncovered silent progressive kidney injury in patients with chronic obstructive pulmonary disease. Kidney Int 2021; 100; 1081-1091.
[7] Roscigno, G., et al.: Urinary Dickkopf-3 and contrast associated kidney damage. J Am CollCardiol 2021; 77: 2667-2676.
[8] Schuster, A., et al.: Dickkopf 3 —A new indicator for the deterioration of allograft function after kidney transplantation. Front Med 2022; doi.org/10.3389/ fmed.2022.885018
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