Combination of an autoantibody panel and alpha-fetoprotein for early detection of hepatitis B virus-associated hepatocellular carcinoma.

The purpose of this study was to identify biomarkers associated with hepatitis B virus-associated hepatocellular carcinoma (HBV-HCC) and to develop a new combination with good diagnostic performance. This study was divided into four phases: discovery, verification, validation and modeling. A total of four candidate tumor-associated autoantibodies (TAAbs) (anti-ZIC2, anti-PCNA, anti-CDC37L1, and anti-DUSP6) were identified by human proteome microarray (52 samples) and bioinformatics analysis. Subsequently, these candidate TAAbs were further confirmed by indirect enzyme-linked immunosorbent assay (ELISA) with two testing cohorts (120 samples for verification and 663 samples for validation). The area under the curve (AUC) for these four TAAbs to identify HBV-HCC patients from chronic hepatitis B (CHB) patients ranged from 0.693-0.739. Finally, a diagnostic panel with three TAAbs (anti-ZIC2, anti-CDC37L1 and anti-DUSP6) was developed. This panel showed superior diagnostic efficiency in identifying early HBV-HCC compared with AFP, with an AUC of 0.834 (95% CI [confidence interval]: 0.772-0.897) for this panel and 0.727 (95% CI: 0.642-0.812) for AFP (P=0.0359). In addition, the AUC for this panel to identify AFP-negative HBV-HCC patients was 0.796 (95% CI: 0.734-0.858), with a sensitivity of 52.4% and a specificity of 89.0%. Importantly, the panel in combination with AFP significantly increased the positive rate for early HBV-HCC to 84.1% (P=0.005) and for late HBV-HCC to 96.3% (P<0.001). Our findings suggest that AFP and the autoantibody panel may be independent but complementary serologic biomarkers for HBV-HCC detection.