The aim of the present study was to determine the presence of HCV RNA in the liver biopsies of patients with abnormal liver tests but without detectable serum HCV RNA and anti-HCV antibodies in sera. Liver biopsies and whole blood of total 31 patients who were negative for anti-HCV antibodies with elevated liver function tests were received at Division of Molecular Diagnostics, University of the Punjab Pakistan from January 2002 to June 2009 for the detection of HCV RNA. HCV RNA status of the subjects was tested by reverse-transcription PCR and quantified using SmartCycler II real-time PCR. HCV genotyping was carried out in HCV RNA positive samples using molecular genotyping method. HCV RNA was found in liver-biopsy specimens from 23 (74.2%) of the total 31 patients negative for anti-HCV antibodies and undetectable serum HCV RNA. HCV RNA of both negative and positive polarity was found in the livers of 8 (25.8%) patients. Genotyping analysis showed that 65% patients were infected with HCV 3a, 17% with 3b, 13% with 1a and 4% patients were found with untypable genotype. In a multivariate logistic regression model, patients having previous history surgeries, male sex and age above 30 years were significantly associated with the presence of occult HCV infection (p<0.05). In conclusion, patients with elevated liver enzymes and negative HCV antibodies and negative serum RNA may have occult HCV infection and its chance increases with previous history of surgeries, in male sex and above 30 years of age.