title: 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.
abstract: BACKGROUND
  Hepatitis C infection recurs after orthotopic liver transplantation for hepatitis C virus (HCV)-related end-stage liver disease. Overlapping histopathologic features may present difficulties in differentiating recurrent HCV in the allograft from other conditions, especially rejection.
  
  
  METHODS
  In this study, we evaluated the presence of HCV-RNA by reverse transcriptase in situ polymerase chain reaction (RT in situ RCR) in hepatic tissue, after orthotopic liver transplantation for HCV-related liver disease. Further, detection of HCV-RNA was correlated with the serum HCV-RNA levels, histopathology, and clinical outcome.
  
  
  RESULTS
  Twenty-five patients were part of this study. Seventeen patients were transplanted for HCV cirrhosis and eight for an underlying disease other than HCV. None of the patients in the non-HCV group had in situ RT-PCR detection of HCV-RNA. Positive RT in situ PCR for HCV was found in 9 of 17 HCV patients, and the patients had a clinical course consistent with recurrent hepatitis C disease. Four of these nine patients had an initial histologic diagnosis of rejection. The other eight patients in the HCV group had negative RT in situ PCR, and none of them had a course compatible with recurrent HCV disease, although four patients were histologically diagnosed as having chronic C hepatitis. The mean HCV-RNA level (log/mL) in the patients who had in situ detection of HCV-RNA was 7.01+/-0.26. Although RT-PCR was negative in 8 of 17 HCV patients, the patients were serologically viremic and the 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 rejection. This may further help in the adjustment of immunosuppression.
authors.affiliation: Department of Surgery, University of Miami School of Medicine, Florida 33136, USA.
authors.forename: G P
authors.initials: GP
authors.lastname: Fragulidis
authors.forename: R E
authors.initials: RE
authors.lastname: Cirocco
authors.forename: D
authors.initials: D
authors.lastname: Weppler
authors.forename: M
authors.initials: M
authors.lastname: Berho
authors.forename: G
authors.initials: G
authors.lastname: Gillian
authors.forename: M
authors.initials: M
authors.lastname: Markou
authors.forename: A
authors.initials: A
authors.lastname: Viciana
authors.forename: V
authors.initials: V
authors.lastname: Esquenazi
authors.forename: J R
authors.initials: JR
authors.lastname: Nery
authors.forename: J
authors.initials: J
authors.lastname: Miller
authors.forename: K R
authors.initials: KR
authors.lastname: Reddy
authors.forename: A G
authors.initials: AG
authors.lastname: Tzakis
mesh_terms: D000328:Adult; D000368:Aged; D001706:Biopsy; D005260:Female; D005784:Gene Amplification; D016174:Hepacivirus / Q000235:genetics*; D006526:Hepatitis C / Q000601:surgery*; D006801:Humans; D008099:Liver / Q000473:pathology*; D016031:Liver Transplantation; D008297:Male; D008875:Middle Aged; D012367:RNA, Viral / Q000097:blood*; D020133:Reverse Transcriptase Polymerase Chain Reaction
chemical_list: D012367:RNA, Viral