Z Gastroenterol 2009; 47 - P5_10
DOI: 10.1055/s-0029-1191972

MALDI-MS analysis reveals reliable results in staging HCV induced fibrogenesis

C Henkel 1, K Schwamborn 1, OA Gressner 2, M Odenthal 3, R Knuechel-Clarke 1, R Weiskirchen 2
  • 1Abteilung für Pathologie, Universitätsklinikum Aachen
  • 2Institut für Klinische Chemie und Pathobiochemie, Universitätsklinikum der RWTH Aachen
  • 3Institute of Pathology, University Hospital Cologne

Introduction

Chronic hepatitis C is characterized by a highly variable clinical course. Approximately 20% of all patients who either do not receive antiviral therapy or fail to respond develop liver cirrhosis within 20 years. Currently, serum markers for hepatic fibrosis diagnosis show a limited sensitivity and specificity so that liver biopsy with high sampling error rate is still used (1). Clinical application of current biomarkers is therefore not feasible. There is an absolute demand on high-throughput methods allowing the reliable assessment of the severity and/or progression of hepatic fibrogenesis.

Methods

We here performed proteomic profiling of serum by matrix-assisted laser desorption ionization time-of-flight (MALDI-TOF) mass spectrometry (MS) in 104 samples of patients with chronic hepatitis B and C that were enrolled by the German Network of Competence for Hepatitis (Hep-Net) and 104 control sera. For prefractionation weak cation exchange surface beads were used (WCX, Bruker Daltonics, Germany). Multi-dimensional analysis of spectra obtained resulted in the definition of a genetic algorithm capable to distinguish class-specific protein expression profiles in serum samples. For analysis ClinProt Tools software (Bruker Daltonics, Germany) was used.

Results

Overall about 114 significant peaks could be detected per single spectrum. The algorithm included 5 of these protein peaks showing p-values <1×10-5 each in the range from 2000 to 6,200 Da for classification on the WCX surface. Using the genetic-algorithm a specificity of 93,2% with a sensitivity of 94,02% could be found. We were also able to differentiate fibrosis stages (1+2 compared to 3+4) with a sensitivity of 83,46%. We conclude that serum profiling is a reliable proteomic technique to identify specific hepatitis-associated expression patterns which are capable to distinguish patients with or without hepatitis (2).

Literatur: (1) Gressner OA, Weiskirchen R, Gressner AM. Biomarkers of hepatic fibrosis, fibrogenesis and genetic pre-disposition pending between fiction and reality. J Cell Mol Med. 2007;11:1031-51. (2) Schwamborn K, Krieg RC, Henkel C, Gressner OA, Wedemeyer I, Odenthal M, Weiskirchen R. Proteomic profiling of serum by MALDI-TOF mass spectrometry is a promising diagnostic technique to analyze the severity and progression of fibrogenesis associated with chronic Hepatitis infection. submitted

HCV - MALDI-MS - fibrogenesis - screening