Acute renal allograft rejection is a serious and treatable complication of renal transplantation. Currently, renal allograft biopsy is required to establish the diagnosis of acute rejection. The purpose of this study was to identify urinary biomarkers of rejection that can accurately establish a diagnosis of acute rejection. We have identified biomarkers that can differentiate the presence or absence of acute renal allograft rejection in a set of patients who received renal allograft biopsy for suspected rejection. Proteins from urine samples collected from 20 patients just prior to biopsy were separated by 2D gel electrophoresis, and gel spot abundance was analyzed to determine differences in patients with rejection, borderline rejection, or no rejection. Artificial neural network analysis correctly identified 5 of 6 patients with rejection (sensitivity: 83%, specificity 86%), 6 of 7 patients with borderline rejection (sensitivity: 86%, specificity 92%), and 6 of 7 patients with no rejection (sensitivity: 86%, specificity 100%). Protein from 16 spots with differential expression between groups was digested and identified using MALDI-TOF mass spectrometry and peptide mass fingerprinting. Proteins identified included alpha-1-microglobulin, zinc-alpha-2 glycoprotein 1 (chain D of this protein was identified in a spot separate from the intact protein), and alpha-1-acid glycoprotein 1 precursor. This analysis is able to successfully differentiate between patients with or without rejection and is the first analysis to identify the differentially expressed proteins in these patients.
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