Real-time reverse transcription polymerase chain reaction detection and quantification of t(1;19) (E2A-PBX1) fusion genes associated with leukaemia

Br J Haematol. 2001 Dec;115(4):826-30. doi: 10.1046/j.1365-2141.2001.03190.x.

Abstract

A real-time reverse transcription polymerase chain reaction (RT-PCR) method is described that enabled the detection and quantification of E2A-PBX1 fusion gene transcripts associated with t(1;19). The method was highly reproducible and offered exceptional sensitivity at 5 fg of fusion transcript per reaction, without the need for a nested PCR primer design. To illustrate the usefulness of this new technology the E2A-PBX1 fusion gene transcript expression level for several human leukaemia cell lines that are positive and negative for cytogenetically detectable t(1;19) was determined. The RCH-ACV had a threefold higher expression of E2A-PBX1 transcripts (600 transcripts per cell) than the other t(1;19) positive 697 (150 transcripts per cell). The only other cell line with detectable E2A-PBX1 was CEM, but the level of expression was < 1 transcript per cell.

Publication types

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

MeSH terms

  • Cell Line
  • Chromosomes, Human, Pair 1*
  • Chromosomes, Human, Pair 19*
  • Computer Systems
  • Gene Expression
  • Homeodomain Proteins / genetics*
  • Humans
  • Leukemia / genetics*
  • Oncogene Proteins, Fusion / genetics*
  • Reverse Transcriptase Polymerase Chain Reaction*
  • Sensitivity and Specificity
  • Translocation, Genetic*

Substances

  • Homeodomain Proteins
  • Oncogene Proteins, Fusion
  • E2A-Pbx1 fusion protein