Toxoplasmosis may present as a severe disease among some Egyptian patients with chronic liver disease (CLD) due to their impaired immune system, changing the course of the disease. The classical diagnosis of toxoplasmosis by serological tests is inadequate for such patients. This study was performed to highlight the role of real-time quantitative PCR (qrtPCR) test in the accurate diagnosis of toxoplasmosis among Egyptian patients with CLD. Seventy patients with CLD and 50 healthy controls were enrolled in this study. All were subjected to full clinical examinations, abdominal ultrasonography, and biochemical analysis of liver enzymes and they were investigated for markers of hepatitis B virus (HBV) and hepatitis C virus (HCV). In addition, Toxoplasma gondii (T. gondii) parasitemia was determined using qrtPCR. The results showed that T. gondii parasitemia was positive in 30% of CLD patients with highly statistically significant (p < 0.001) compared with the control group (6%). Co-infection in both T. gondii/HBV and T. gondii/HCV was 33.3% and 31.4%, respectively, with a highly significant association between T. gondii parasitemia and HCV viral load. Moreover, the results showed a significant increase of liver enzymes in the serum of patients positive for T. gondii compared with negative patients. An association between T. gondii infection and CLD was observed, and further studies will be needed to define the mechanism of this association.