Detection and direct sequence identification of BCR-ABL mRNA in Ph+ chronic myeloid leukemia

Exp Hematol. 1993 Dec;21(13):1719-24.

Abstract

The reverse transcriptase-polymerase chain reaction (RT-PCR) for BCR-ABL mRNA is increasingly used to diagnose and monitor patients with Ph+ chronic myeloid leukemia (CML). We investigated an alternative approach to detect BCR-ABL mRNA in CML in order to overcome some of the potential drawbacks of RT-PCR. Nucleic acid sequence based amplification (NASBA) is a homogeneous, isothermal, in vitro process that provides the direct amplification of RNA. Peripheral blood from seven patients with Ph+ CML and Ph+ EM-2 cells were investigated by NASBA and RT-PCR. A nested set of four primers flanking the BCR-ABL junction was used in two serial NASBA reactions performed for 2 hours. The two methods were fully concordant for detection of transcripts with bcr3-abl2 and bcr2-abl2 junctions. Ethidium bromide fluorescence with NASBA indicated in repeated experiments that similar quantities of total RNA from patient material contained different amounts of BCR-ABL mRNA. The data suggest that direct amplification of RNA is suitable for identifying and monitoring patients with Ph+ CML and may provide a means to quantify BCR-ABL mRNA levels.

Publication types

  • Comparative Study

MeSH terms

  • Base Sequence
  • Fusion Proteins, bcr-abl / genetics*
  • Humans
  • Leukemia, Myelogenous, Chronic, BCR-ABL Positive / diagnosis
  • Leukemia, Myelogenous, Chronic, BCR-ABL Positive / genetics*
  • Molecular Sequence Data
  • Nucleic Acid Amplification Techniques*
  • Nucleic Acid Hybridization
  • Polymerase Chain Reaction
  • RNA, Messenger / blood*
  • RNA, Messenger / chemistry
  • Sequence Analysis, RNA

Substances

  • RNA, Messenger
  • Fusion Proteins, bcr-abl