In situ polymerase chain reaction amplification of HIV-1 DNA in brain tissue

P M Strappe, T H Wang, C A McKenzie, S Lowrie, P Simmonds, J E Bell

Research output: Contribution to journalArticlepeer-review

Abstract

A direct in situ polymerase chain reaction (IS-PCR) assay is described for the detection of HIV-1 proviral DNA in formalin fixed paraffin embedded brain tissue. Biotin-16-dUTP is incorporated during the PCR process and microwave pretreatment of tissue sections ensures that no non-specific incorporation into damaged or nicked genomic DNA occurs. Two methods are compared to detect the biotinylated amplified product, the use of an avidin-biotin-alkaline phosphatase complex (ABC) and the application of tyramide signal amplification (TSA) which allows both chromogenic and fluorescence detection. TSA detection enhances the sensitivity of IS-PCR, permitting fewer PCR cycles and preserving tissue morphology.
Original languageEnglish
Pages (from-to)119-27
Number of pages9
JournalJournal of Virological Methods
Volume70
Issue number2
DOIs
Publication statusPublished - Feb 1998

Keywords

  • Brain
  • DNA, Viral
  • Formaldehyde
  • HIV-1
  • Humans
  • Microwaves
  • Paraffin Embedding
  • Polymerase Chain Reaction
  • Sensitivity and Specificity
  • Tissue Fixation

Fingerprint

Dive into the research topics of 'In situ polymerase chain reaction amplification of HIV-1 DNA in brain tissue'. Together they form a unique fingerprint.

Cite this