High quality of DNA retrieved for Southern blot hybridization from microwave-fixed, paraffin-embedded liver tissues.

Author(s): Hsu HC, Peng SY, Shun CT

Publication: J Virol Methods, 1991, Vol. 31, Page 251-61

PubMed ID: 1864908 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 fixative type and delivery method on IHC detection of Hepatitis B surface antigen (HBSAg) and epitopes preS1 and preS2 in liver specimens.

   Summary of Findings:

   HBSAg and the preS1 epitope were detected regardless of fixation method, but detection of the preS2 epitope decreased when formalin fixation was used and was most intense when fixation was by microwave-accelerated ethanol fixation or microwave fixation (tissue in PBS).

   Biospecimens

   • Tissue - Liver

   Preservative Types

   • Formalin
   • Frozen
   • Other Preservative
   • Ethanol

   Diagnoses:

   • Neoplastic - Carcinoma

   Platform:

   Analyte	Technology Platform
   Protein	Immunohistochemistry

   Pre-analytical Factors:

   Classification	Pre-analytical Factor	Value(s)
   Biospecimen Preservation	Type of fixation/preservation	Alcohol Ethanol Formalin (buffered) Snap frozen
   Biospecimen Preservation	Method of fixative delivery	Immersion Microwaved
   Immunohistochemistry Specific	Targeted peptide/protein	Hepatitis B surface antigen Pre S1 epitope of hepatitis B Pre S2 epitope of hepatitis B

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2. Study Purpose

   The purpose of this study was to determine the effects of fixative type and delivery method on Southern blot detection of HBV and procollagen and PCR amplification of HBV core gene.

   Summary of Findings:

   Immersion or microwave-accelerated ethanol fixation and straight microwave fixation (specimens in PBS) resulted in 80% of the DNA being spoolable. In addition, the DNA appeared as a smear larger than 23 kb on agarose gels and southern hybridization patterns for procollagen and HBV matched those from frozen tissue. However, when tissues were formalin-fixed by microwave, only 40% of the DNA was spoolable, the DNA mobility was hindered, and the procollagen and HBV signals were weaker. When tissues were formalin-fixed by immersion, there was no spoolable DNA, little or no hybridization signal, and reduced PCR amplification.

   Biospecimens

   • Tissue - Liver

   Preservative Types

   • Formalin
   • Frozen
   • Ethanol

   Diagnoses:

   • Hepatitis
   • Neoplastic - Carcinoma

   Platform:

   Analyte	Technology Platform
   DNA	Southern blot
   DNA	PCR
   DNA	Macroscopic observation

   Pre-analytical Factors:

   Classification	Pre-analytical Factor	Value(s)
   Biospecimen Preservation	Type of fixation/preservation	Ethanol Formalin (buffered) Snap frozen
   Biospecimen Preservation	Method of fixative delivery	Immersion Microwaved
   PCR Specific	Targeted nucleic acid	Hepatitis B virus core
   Southern blot Specific	Targeted nucleic acid	Hepatitis B virus Procollagen

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