Early detection of HCC increases the potential for curative treatment and improves survival. To facilitate early detection of HCC, this study sought to identify novel diagnostic markers of HCC using surface-enhanced laser desorption ionization time-of-flight mass spectrometry (SELDI-TOF/MS) ProteinChip technology. Serum samples were obtained from 153 patients with or without HCC, all of whom had been diagnosed with HCV-associated chronic liver disease. To identify proteins associated with HCC, serum samples were analyzed using SELDI-TOF/MS. We constructed an initial decision tree for the correct diagnosis of HCC using serum samples from patients with (n ؍ 35) and without (n ؍ 44) HCC. Six protein peaks were selected to construct a decision tree using this first group. The efficacy of the decision tree was then assessed using a second group of patients with (n ؍ 29) and without (n ؍ 33) HCC. The sensitivity and specificity of this decision tree for the diagnosis of HCC were 83% and 76%, respectively. For a third group, we analyzed sera from seven patients with HCC obtained before the diagnosis of HCC by ultrasonography (US) and from five patients free of HCC for the past 3 years. Use of these diagnostic markers predicted the diagnosis of HCC in six of these seven patients before HCC was clinically apparent without any false positives. Conclusion: Serum profiling using the SELDI ProteinChip system is useful for the early detection and prediction of HCC in patients with chronic HCV infection. (HEPATOLOGY 2007;45:948-956.) A pproximately 170 million people worldwide are infected with HCV, which when persistent can progress to HCC. The incidence of HCC is rising; in the United States over the past 2 decades, age-specific incidence has shifted toward younger people. 1 IFN or combined IFN and ribavirin, which are currently the only effective treatments for chronic hepatitis C, reduce the occurrence of HCC. 2,3 Some patients, however, do not receive IFN treatment or fail to clear HCV even with IFN treatment. In addition, a subset of individuals remain unaware that they are infected with HCV; in these patients, HCC may present only in the advanced stage. The prognosis of patients presenting with symptoms related to HCC is extremely poor. In contrast, early detection of HCC before the onset of clinical symptoms can lead to curative treatment, significantly improving prognosis.Several methods developed for the diagnosis of HCC, including evaluation of serum markers, ultrasonography (US), computed tomography (CT), and magnetic resonance imaging, have been tested clinically. Alpha-fetoprotein (AFP) and des-gamma carboxy prothrombin (DCP), serum proteins that are elevated in HCC, have been the most widely used markers. Although routine screening offers the best chance for early tumor detection and improved survival, the reported sensitivities and specificities of elevated serum AFP and DCP levels vary significantly. [4][5][6][7][8][9] In addition, AFP levels are elevated in only 30% to 40% of patients with HCC, par...