The aim of this study was to compare the results of nine non-invasive serum biomarkers with liver biopsies to predict liver fibrosis stage. HCV-RNA-positive, HCV genotype 1, treatment-naive patients with chronic HCV infections were included from 14 centers (n = 77). The platelet count, AST/ALT ratio (AAR), cirrhosis discriminate score (CDS), FIB4, AST/platelet ratio index (APRI), age-platelet (AP) index, Goteborg University cirrhosis index (GUCI), FibroTest, and ActiTest were calculated and compared to histologic findings. All serum biomarkers, except AAR, were weakly or moderately correlated with liver biopsy results (ISHAK fibrosis score). The mean scores of FibroTest, FIB4, APRI, and AP index were significantly different between F0-F2 and F3-F4 groups and the negative predictive values (NPVs) of the F3-F4 group were 95%, 85%, 85%, and 83%, respectively, for these serum biomarkers. Our study suggests that serum biomarkers may help to diagnose significant fibrosis but inadequate to detect fibrosis in early stages. Although liver biopsy is still the gold standard to diagnose liver fibrosis, FibroTest, FIB4, APRI, or AP index may be used to exclude significant fibrosis with > 80% NPV.