Detection of BCR/ABL transcripts in chronic myeloid leukaemia by polymerase chain reaction and DNA enzyme immunoassay: a DNA probe assay without DNA labelling

Br J Haematol. 1993 Feb;83(2):334-9. doi: 10.1111/j.1365-2141.1993.tb08291.x.

Abstract

The detection of the t(9;22) translocation, typical of chronic myeloid leukaemia (CML), can be accomplished by cytogenetical detection of Philadelphia (Ph1) chromosome or by molecular analysis of the bcr/abl fusion gene with nucleic acid probes after amplification by polymerase chain reaction (PCR). PCR-based approaches are now widely used for follow up of CML patients during therapy or after bone marrow transplantation (BMT). We describe here a microtitre, colorimetric assay (DNA Enzyme Immunoassay, DEIA) for analysis of t(9;22) translocation after enzymatical amplification of RNA from CML patients. This assay is based on the use of a monoclonal antibody specifically reacting with double stranded DNA, i.e. with hybridized DNA. The assay represents a nonisotopic alternative to other current hybridization assays and requires no modifications of primers, probe or target DNA.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adult
  • Base Sequence
  • Chromosomes, Human, Pair 22
  • Chromosomes, Human, Pair 9
  • DNA Probes
  • Female
  • Fusion Proteins, bcr-abl / analysis*
  • Humans
  • Immunoblotting
  • Immunoenzyme Techniques*
  • Leukemia, Myelogenous, Chronic, BCR-ABL Positive / genetics*
  • Male
  • Middle Aged
  • Molecular Sequence Data
  • Oligonucleotide Probes
  • Philadelphia Chromosome*
  • Polymerase Chain Reaction
  • RNA, Messenger / analysis
  • Transcription, Genetic
  • Translocation, Genetic

Substances

  • DNA Probes
  • Oligonucleotide Probes
  • RNA, Messenger
  • Fusion Proteins, bcr-abl