Comparative studies on the detection of hepatitis B virus DNA in frozen and paraffin sections by the polymerase chain reaction.

Author(s): Chen ML, Shieh YS, Shim KS, Gerber MA

Publication: Mod Pathol, 1991, Vol. 4, Page 555-8

PubMed ID: 1758869 PubMed Review Paper? No

Purpose of Paper

Conclusion of Paper

Studies

1. Study Purpose

   The purpose of this study was to compare DNA extracted from frozen and FFPE liver specimens by two methods for detection of HBV by PCR.

   Summary of Findings:

   Chronic active HBV infection and cirrhosis were confirmed by H&E staining and immunohistochemistry. Approximately 2 times more DNA was obtained using the PS method with frozen specimens than with FFPE specimens. The DNA yield from the PS method was similar to the quantity obtained using the CS method for both FFPE and frozen specimens. The most efficient DNA amplification was observed using PS extracted DNA from frozen specimens, while the least efficient amplification was observed using CS extracted DNA from frozen specimens. In FFPE specimens, the CS method resulted in approximately 300% greater amplification than if the DNA was extracted by the PS method. No differences between extraction methods were observed with different primer pairs.

   Biospecimens

   • Tissue - Liver

   Preservative Types

   • Formalin
   • Frozen

   Diagnoses:

   • Cirrhosis
   • Hepatitis
   • Normal

   Platform:

   Analyte	Technology Platform
   DNA	PCR
   Morphology	H-and-E microscopy
   Protein	Immunohistochemistry

   Pre-analytical Factors:

   Classification	Pre-analytical Factor	Value(s)
   Biospecimen Preservation	Type of fixation/preservation	Formalin (buffered) Frozen
   Analyte Extraction and Purification	Analyte isolation method	Proteinase K -SDS followed by phenol chloroform extraction Boiling in water with chelex-100 followed by sephadex G-50 mini-column chromatography
   PCR Specific	Targeted nucleic acid	Hepatitis B S gene Hepatitis B C gene Hepatitis B X gene

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