Discrimination of closely homologous HPV types by nonisotopic in situ hybridization: definition and derivation of tissue melting temperatures

C S Herrington, A K Graham, D M Flannery, J Burns, J O McGee

Research output: Contribution to journalArticlepeer-review

Abstract

It is generally assumed that nucleic acid association during in situ hybridization reactions is similar to that of nucleic acid association in solution. This assumption has been investigated by detecting closely homologous human papillomavirus types 6 and 11 by in situ hybridization as a model for the evaluation of stringency conditions in clinical biopsies. By examining matched and mismatched, labelled and target sequences under various stringency conditions, empirical DNA-DNA stability curves and their derivative equations for tissue melting temperatures (Tmt) were derived. The corresponding values for Tmt are 10-20 degrees C higher than their solution equivalents. These data, supported by polymerase chain reaction experiments, demonstrate that closely homologous viral DNAs cross linked in tissue by formaldehyde fixation do not interact with the corresponding labelled probes as predicted from solution kinetic equations. This not only has theoretical implications but is also relevant to the accuracy of clinical diagnostic testing.

Original languageEnglish
Pages (from-to)545-54
Number of pages10
JournalHistochemical Journal
Volume22
Issue number10
Publication statusPublished - Oct 1990

Keywords

  • Base Sequence
  • Biopsy
  • Condylomata Acuminata
  • DNA, Viral
  • Humans
  • Molecular Sequence Data
  • Nucleic Acid Hybridization
  • Papillomaviridae
  • Polymerase Chain Reaction
  • Sequence Homology, Nucleic Acid
  • Temperature
  • Tumor Virus Infections

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