Detection of hepatitis C virus RNA sequences by polymerase chain reaction in fixed liver tissue.

Author(s): Akyol G, Dash S, Shieh YS, Malter JS, Gerber MA

Publication: Mod Pathol, 1992, Vol. 5, Page 501-4

PubMed ID: 1344813 PubMed Review Paper? No

Purpose of Paper

Conclusion of Paper

Studies

1. Study Purpose

   The purpose of this study was to determine the effects of formalin fixation duration, amplicon length, and amplicon location on HCV detection in FFPE and frozen liver specimens.

   Summary of Findings:

   The NT, core and NS3 regions of HCV were amplifiable in frozen specimens and could be detected in as little as 1 ng of RNA, but only the NT region was amplifiable in FFPE specimens and 1 ug of RNA was necessary. Use of different primers that amplified a longer fragment of the NT region allowed for amplification of all 3 HCV positive specimens, but primers that amplified a smaller region only detected HCV in the 2 chronic cases. Amplification of the longer NT amplicon was preserved in specimens fixed for 6-48 h, but was lost in specimen fixed for 1 week even though a control fragment of albumin was still detected. Importantly, HCV NT fragments were not detected in specimens without hepatitis C.

   Biospecimens

   • Tissue - Liver

   Preservative Types

   • Formalin
   • Frozen

   Diagnoses:

   • Hepatitis
   • Not specified
   • Cardiovascular Disease
   • Neoplastic - Carcinoma
   • Cirrhosis

   Platform:

   Analyte	Technology Platform
   RNA	RT-PCR

   Pre-analytical Factors:

   Classification	Pre-analytical Factor	Value(s)
   Biospecimen Preservation	Type of fixation/preservation	Formalin (buffered) Snap frozen
   Biospecimen Preservation	Time in fixative	6 h 24 h 48 h 1 week
   RT-PCR Specific	Targeted nucleic acid	Noncoding region Structural core Nonstructural Albumin
   RT-PCR Specific	Length of gene fragment	589/338 bp 219/142 bp 162/115 bp 404/185 bp
   Preaquisition	Diagnosis/ patient condition	HCV positive HCV negative

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