Application of the hybridization AT-tailing method for detection of human immunodeficiency virus RNA in cells and simian immunodeficiency virus RNA in formalin-fixed and paraffin-embedded tissues

J Virol Methods. 1999 Aug;81(1-2):169-77. doi: 10.1016/s0166-0934(99)00072-5.

Abstract

An in situ hybridization (ISH) AT-tailing method (HybrAT) was developed for the detection of viral genomes in infected cells and tissues. The method consists of hybridization with oligonucleotide probe which has a 3' end oligo d(A-T) tag, followed by elongation of the oligo d(A-T) by deltaTth DNA polymerase in the presence of the labeled nucleotide. The in situ HybrAT detected human immunodeficiency virus type 1 (HIV-1) in cells and simian immunodeficiency virus (SIV) in formalin-fixed and paraffin embedded sections with a sensitivity comparable to RNA ISH. The advantage of this method over other methods is discussed.

MeSH terms

  • Animals
  • Cell Line
  • Formaldehyde
  • HIV-1 / genetics*
  • HIV-1 / isolation & purification
  • Humans
  • In Situ Hybridization / methods*
  • Macaca mulatta
  • Male
  • Oligodeoxyribonucleotides / genetics*
  • Oligonucleotide Probes / genetics*
  • Paraffin Embedding
  • Poly dA-dT / genetics*
  • RNA, Messenger / isolation & purification
  • RNA, Viral / isolation & purification*
  • Reagent Kits, Diagnostic
  • Simian Immunodeficiency Virus / genetics*
  • Simian Immunodeficiency Virus / isolation & purification
  • Tissue Fixation

Substances

  • Oligodeoxyribonucleotides
  • Oligonucleotide Probes
  • RNA, Messenger
  • RNA, Viral
  • Reagent Kits, Diagnostic
  • oligo (dT) poly (A)
  • Formaldehyde
  • Poly dA-dT