Real-time quantitative assay for routine testing of HCV RNA in formalin-fixed, paraffin-embedded liver samples.
The assessment of hepatitis C virus (HCV) RNA in liver tissues is clinically relevant in cases where histology, liver function tests, and HCV serology are not sufficient for a definitive diagnosis of HCV-related hepatitis. We analyzed 215 formalin-fixed, paraffin-embedded liver needle biopsies from patients infected with HCV genotypes 1b and 2. HCV RNA extracted from paraffin sections were quantified by means of a TaqMan real-time reverse transcription-polymerase chain reaction method. The quantification of HCV RNA in liver tissue was correlated with the amount of HCV detected by immunohistochemistry (IHC) on paired frozen biopsies, the HCV RNA load in the serum, and the main serum tests of liver function and cholestasis. HCV RNA was detected by real-time reverse transcription-polymerase chain reaction in 169 liver biopsies (78.6%) with a mean value of 13.59+/-37.25 IU/ng. Tissue HCV RNA levels strongly correlated with the IHC results (P<0.001, Spearman test), HCV serum load (P<0.001), aspartate aminotransferase (P=0.001), gamma-glutamyl transpeptidase (P=0.012), and aspartate aminotransferase/alanine aminotransferase ratio (P=0.029). HCV RNA was amplified in up to 7-year-old archival tissue samples. Real-time HCV RNA quantification on archival liver tissue may be clinically relevant in case of "occult" HCV infection or for the diagnosis of patients with known HCV infection and hepatic dysfunction but seronegative for HCV RNA. The assessment of the levels of HCV RNA in the liver might also be important for monitoring the effectiveness of antiviral therapy and the progression of disease in patients with chronic HCV hepatitis.
Molecular and Transplantation Pathology Laboratory, F. Addarii Institute of Oncology, University of Bologna, Italy.
Elisa
E
Gruppioni
Francesco
F
Vasuri
Michelangelo
M
Fiorentino
Elisa
E
Capizzi
Annalisa
A
Altimari
Maria G
MG
Pirini
Gian L
GL
Grazi
Deborah
D
Malvi
Walter F
WF
Grigioni
Antonia
A
D'Errico-Grigioni
D005557:Formaldehyde; D016174:Hepacivirus / Q000235:genetics / Q000302:isolation & purification*; D006526:Hepatitis C / Q000097:blood / Q000175:diagnosis / Q000821:virology*; D006801:Humans; D007124:Immunoenzyme Techniques; D008099:Liver / Q000473:pathology / Q000821:virology*; D008111:Liver Function Tests; D025202:Molecular Diagnostic Techniques; D016612:Paraffin Embedding; D011237:Predictive Value of Tests; D012367:RNA, Viral / Q000032:analysis* / Q000235:genetics; D015203:Reproducibility of Results; D012189:Retrospective Studies; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D016707:Tissue Fixation
D012367:RNA, Viral; D005557:Formaldehyde