In situ enzymatic oligonucleotide amplification of hepatitis C virus RNA in liver biopsy specimens (reverse transcriptase in situ polymerase chain reaction) after orthotopic liver transplantation for hepatitis C-related liver disease

Background. Hepatitis C infection recurs after orthotopic liver transplanta tion for hepatitis C virus (HCV)-related end-stage liver disease. Overlappi ng histopathologic features may present difficulties in differentiating rec urrent HCV in the allograft from other conditions, especially rejection. Methods, In this study, we evaluated the presence of HCV-RNA by reverse tra nscriptase in situ polymerase chain reaction (RT in situ RCR) in hepatic ti ssue, after orthotopic liver transplantation for I-ICV-related liver diseas e. Further, detection of HCV-RNA was correlated with the serum HCV-RNA leve ls, histopathology, and clinical outcome. Results, Twenty-five patients were part of this study. Seventeen patients w ere transplanted for HCV cirrhosis and eight for an underlying disease othe r than HCV. None of the patients in the non-HCV group had in situ RT-PCR de tection of HCV-RNA. Positive RT in situ PCR for HCV was found in 9 of 17 HC V patients, and the patients had a clinical course consistent with recurren t hepatitis C disease. Four of these nine patients had an initial histologi c diagnosis of rejection. The other eight patients in the HCV group had neg ative RT in situ PCR, and none of them had a course compatible with recurre nt HCV disease, although four patients were histologically diagnosed as hav ing chronic C hepatitis. The mean HCV-RNAlevel (log/mL) in the patients who had in situ detection of HCV-RNA was 7.01+/-0.26. Although RT-PCR was nega tive in 8 of 17 HCV patients, the patients were serologically viremic and t he mean HCV-RNA level was 6.05 +/- 0.33 (P=0.03). Conclusions. Our findings indicate that the HCV in situ RT-PCR assay may be helpful in the differentiation of recurrent hepatitis C disease from rejec tion. This may further help in the adjustment of immunosuppression.