Real-time quantitative assay for routine testing of HCV RNA in formalin-fixed, paraffin-embedded liver samples.

Author(s): Gruppioni E, Vasuri F, Fiorentino M, Capizzi E, Altimari A, Pirini MG, Grazi GL, Malvi D, Grigioni WF, D'Errico-Grigioni A

Publication: Diagn Mol Pathol, 2009, Vol. 18, Page 232-8

PubMed ID: 19861893 PubMed Review Paper? No

Purpose of Paper

Conclusion of Paper

Studies

1. Study Purpose

   The purpose of this study was to compare real-time PCR of FFPE liver specimens with IHC, serum viral load, and liver function test results for the diagnosis of hepatitis C.

   Summary of Findings:

   HCV was detected by real-time PCR in 79% of the 215 cases studied including 8 cases found to be negative by IHC and in one case that had been archived for 7 years. No cases were found to be positive by IHC and negative by real-time PCR, indicating that real-time PCR is more sensitive then IHC for HCV detection. Strong correlations were observed between positive real-time PCR results and IHC staining (p<0.001), serum viral load (p<0.001), gamma-glutamyl transpeptidase activity (p=0.012), aspartate aminotransferase activity (AST) (p=0.001), and AST alanine aminotransferase ratio (p=0.029). In contrast, IHC results only correlated with AST activity (p=0.025) and serum viral load did not correlate significantly with any other serological marker. When HCV infection was grouped based on percentage of hepatocytes positive for HCV by IHC, real-time PCR (p<0.001), serum viral load (p=0.001) and AST levels (p=0.014) were significantly different among the groups. In conclusion, real-time PCR was found to be more sensitive then IHC at detecting HCV in FFPE specimens and correlated with IHC results, serum viral load and several liver function tests.

   Biospecimens

   • Tissue - Liver
   • Bodily Fluid - Serum

   Preservative Types

   • Formalin

   Diagnoses:

   • Hepatitis

   Platform:

   Analyte	Technology Platform
   RNA	Real-time qRT-PCR
   Protein	Immunohistochemistry
   Protein	Clinical chemistry/auto analyzer
   Protein	Enzyme assay
   Small molecule	Clinical chemistry/auto analyzer

   Pre-analytical Factors:

   Classification	Pre-analytical Factor	Value(s)
   Real-time qRT-PCR Specific	Technology platform	Immunohistochemistry Serum viral load Liver function tests
   Biospecimen Acquisition	Biospecimen location	Serum Liver
   Real-time qRT-PCR Specific	Targeted nucleic acid	GUS HCV

   View more