Comparison of formalin, ethanol, and Histochoice fixation on the PCR amplification from paraffin-embedded breast cancer tissue.

Author(s): Giannella C, Zito FA, Colonna F, Paradiso A, Marzullo F, Alaibac M, Schittulli F

Publication: Eur J Clin Chem Clin Biochem, 1997, Vol. 35, Page 633-5

PubMed ID: 9298355 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, fixation duration, and amplicon size on DNA quality and PCR success using paraffin embedded breast carcinoma specimens.

   Summary of Findings:

   The quality of the DNA was good when the specimens were fixed in ethanol, but histochoice fixation resulted in partially degraded DNA, and formalin fixation resulted in severely degraded DNA and decreased yield. There was no clear effect of fixation duration on DNA degradation. Amplification of a 280 and a 530 bp product was possible in all ethanol-fixed specimens, and in all specimens fixed in histochoice for 24 h. Amplification of the 530 bp product was not possible in 1 of 6 specimens fixed in histochoice for 72 h or in formalin for 24 h. After fixation in formalin for 72 h, the 280 bp product failed to amplify in 1 of 6 specimens and the 530 bp product failed to amplify in 3 of 6 specimens.

   Biospecimens

   • Tissue - Breast

   Preservative Types

   • Formalin
   • Other Preservative
   • Ethanol

   Diagnoses:

   • Neoplastic - Carcinoma

   Platform:

   Analyte	Technology Platform
   DNA	PCR
   DNA	Electrophoresis
   DNA	Spectrophotometry

   Pre-analytical Factors:

   Classification	Pre-analytical Factor	Value(s)
   Biospecimen Preservation	Type of fixation/preservation	Ethanol Formalin (buffered) Histochoice
   Biospecimen Preservation	Time in fixative	24 h 72 h
   PCR Specific	Length of gene fragment	280 bp 530 bp
   PCR Specific	Targeted nucleic acid	Androgen receptor Phosphoglycerokinase

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