Variability in beta-globin and HPV DNA amplification by PCR from fixed tissues

Mod Pathol. 1991 Sep;4(5):667-70.

Abstract

Paraffin-embedded tissues from a variety of sources and treated with different fixatives were tested for beta-globin and HPV amplification by polymerase chain reaction (PCR). In tests of tissues collected in the previous 2 to 3 yr, excellent rates (87% to 93%) for beta-globin amplification were obtained for specimens fixed in buffered formalin, Bouin's, and Hartmann's solutions. In contrast, the rate of beta-globin amplification was low for tissues fixed in Hollande's solution (7%) and in Hartmann's solution with eosin (33%). The results of beta-globin amplification from archival tissues stored for variable time periods showed no decrease in the amplification rate with longer periods of storage. Human papillomaviruses (HPVs) were identified in 17% of globin negative and in 43% of globin positive tissues. HPV-16 amplification was more efficient when the targeted DNA sequence was small. Variability in amplification depends not only on the type of fixative used, but also on other ill-defined factors. Therefore, conditions for optimal amplification should be determined before undertaking studies of archival material.

Publication types

  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Blotting, Southern
  • Cervix Uteri / chemistry
  • Cervix Uteri / microbiology
  • DNA, Viral / analysis
  • DNA, Viral / genetics*
  • Female
  • Gene Amplification / genetics*
  • Genetic Variation / genetics
  • Globins / analysis
  • Globins / genetics*
  • Humans
  • Papillomaviridae / genetics*
  • Papillomaviridae / isolation & purification
  • Polymerase Chain Reaction
  • Vulva / chemistry
  • Vulva / microbiology

Substances

  • DNA, Viral
  • Globins