A rapid practical RT-PCR-based approach for the detection of the PML/RAR alpha fusion transcript in acute promyelocytic leukemia

Am J Clin Pathol. 1999 Aug;112(2):256-62. doi: 10.1093/ajcp/112.2.256.

Abstract

The t(15;17) and its molecular equivalent, PML/RAR alpha gene fusion, is strongly associated with acute promyelocytic leukemia (APL). Since treatment response to all-trans retinoic acid correlates directly with PML/RAR alpha, expeditious documentation is critical to patient care. We have designed an extremely rapid, practical, polymerase chain reaction (PCR)-based method using a rapid air thermal cycler to detect type A, B, and B-variant fusion patterns of PML/RAR alpha. We examined 15 cases of APL and 13 cases of leukemias other than APL with a nested reverse-transcription PCR assay. Three APL samples were type A, 11 were type B, and 1 was a B variant based on gel band patterns. PCR products exhibited positive probe hybridization signals and had sequences containing type A, B, or B-variant fusion patterns. PCR amplification of PML/RAR alpha was complete in 22 minutes, and the entire test required 4 1/2 hours. This method permits exceptional turnaround time and is an alternative to cytogenetics and slower PCR assays.

Publication types

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

MeSH terms

  • Artificial Gene Fusion / methods*
  • Base Sequence
  • Chromosomes, Human, Pair 15
  • Chromosomes, Human, Pair 17
  • DNA Primers / chemistry
  • DNA, Neoplasm / analysis
  • Humans
  • Leukemia, Promyelocytic, Acute / genetics*
  • Leukemia, Promyelocytic, Acute / pathology
  • Molecular Sequence Data
  • Neoplasm Proteins / genetics*
  • Nuclear Proteins*
  • Oncogene Proteins, Fusion / genetics*
  • Promyelocytic Leukemia Protein
  • Reverse Transcriptase Polymerase Chain Reaction / methods*
  • Transcription Factors / genetics*
  • Translocation, Genetic
  • Tumor Suppressor Proteins

Substances

  • DNA Primers
  • DNA, Neoplasm
  • Neoplasm Proteins
  • Nuclear Proteins
  • Oncogene Proteins, Fusion
  • Promyelocytic Leukemia Protein
  • Transcription Factors
  • Tumor Suppressor Proteins
  • PML protein, human