Objective To systematically review the diagnostic value of anti-HCV in serum tested by the third-generation enzyme-linked immunosorbent assay (ELISA 3) in patients with hepatitis C. Methods Such databases as PubMed, MEDLINE, EMbase, The Cochrane Library (Issue 1, 2013), EBSCO, CBM, CNKI, VIP and WanFang Data were electronically and comprehensively searched for relevant studies on the diagnostic value of anti-HCV in serum tested by ELISA 3 in patients with hepatitis C from inception to January 1st, 2013. Relevant journals were also manually retrieved. QUADAS items were used to evaluate the quality of the included studies. Then meta-analysis was performed using Meta-Disc 1.4 software to calculate pooled effect size in sensitivity (SEN), specificity (SPE), likelihood ratio (±LR), diagnostic odds ratio (DOR) and summary receiver operating characteristic curve (SROC curve), and area under the curve (AUC) as well. Results Finally, nine studies were included, involving 1 182 patients with hepatitis C diagnosed by the gold standard and 4 764 patients with non-hepatitis C diseases. The results of meta-analysis using random model showed (SEN=0.96, 95%CI 0.95 to 0.97; SPE=0.96, 95%CI 0.96 to 0.97; +LR=42.21, 95%CI 10.23 to 174.23; –LR=0.02, 95%CI 0.01 to 0.09; DOR=1 635.89, 95%CI 372.37 to 7 186.83; AUC=0.996 5). Conclusion Anti-HCV in serum tested by ELISA 3 has fairly high value in the diagnosis of patients with hepatitis C.