Quantitative RT-PCR of PML-RARα Transcripts in Acute Promyelocytic Leukemia

  • Harry Iland
  • Francisca Springall
  • Tao Zeng
  • Katrina Bradfield


Detection of PML-RARα fusion transcripts in acute promyelocytic leukemia (APL) by RT-PCR is a useful adjunct to cytogenetic analysis at the time of diagnosis, and is the method of choice for detection of minimal residual disease. Since PML-RARα transcripts are frequently detectable after successful remission induction with all-trans retinoic acid (ATRA), we have established a competitive RT-PCR protocol for quantitation of PML-RARα transcripts which could be used to assess the effects of ATRA induction and subsequent chemotherapy consolidation. In addition to quantitation of the target transcript (PML-RARα), simultaneous quantitation of a second template (transferrin receptor transcripts) provides a control for the quality of RNA extraction and for the efficiency of cDNA synthesis, since the level of PML-RARα transcripts can be expressed relative to the level of transferrin receptor transcripts. For patients with APL, we anticipate that decisions regarding the requirement for additional chemotherapy, or the optimal timing of allogeneic transplantion, will be facilitated by the ability to detect progressive increases in fusion transcripts.


Chronic Myeloid Leukemia Acute Promyelocytic Leukemia Minimal Residual Disease Transferrin Receptor Fusion Transcript 
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  1. 1.
    . Rabbitts TH. Chromosomal translocations in human cancer. Nature 372: 143, 1994.PubMedCrossRefGoogle Scholar
  2. 2.
    . Warrell RP Jr, de Thé H, Wang Z-Y, Degos L. Acute promyelocytic leukemia. New Engl J Med 329: 177, 1993.PubMedCrossRefGoogle Scholar
  3. 3.
    . Zelent A. Translocation of the RARα locus to the PML or PLZF gene in acute promyelocytic leukaemia. Brit J Haematol 86: 451, 1994.CrossRefGoogle Scholar
  4. 4.
    . Dhingra K, Kurzrock R, Kantarjian H, Baine R, Eastman PS, Ku S, Gutterman JU, Talpaz M. Minimal residual disease in interferon-treated chronic myelogenous leukemia: results and pitfalls of analysis based on polymerase chain reaction. Leukemia 6: 754, 1992.PubMedGoogle Scholar
  5. 5.
    . Miyamura K, Barrett J, Kodera Y, Saito H. Minimal residual disease after bone marrow transplantation for chronic myelogenous leukemia and implications for graft-versus-leukemia effect: a review of recent results. Bone Marrow Transplantation 14: 201, 1994.PubMedGoogle Scholar
  6. 6.
    . Miller WH Jr, Kakizuka A, Frankel SR, Warrell RP Jr, DeBlasio A, Levine K, Evans RM, Dmitrovsky E. Reverse transcription polymerase chain reaction for the rearranged retinoic acid receptor α clarifies diagnosis and detects minimal residual disease in acute promyelocytic leukemia. Proc Natl Acad Sci (USA) 89: 2694, 1992.CrossRefGoogle Scholar
  7. 7.
    . Cross NCP. Quantitative PCR techniques and applications. Brit J Haematol 89: 693, 1995.CrossRefGoogle Scholar
  8. 8.
    . Chomczinski P and Sacchi N. Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Analytical Biochem 162: 156, 1987.Google Scholar
  9. 9.
    . Morgan GJ, Shiach C, Potter M. The clinical value of detecting gene rearrangements in acute leukaemias. Brit J Haematol 88: 459, 1994.CrossRefGoogle Scholar
  10. 10.
    . Kohler S, Galili N, Sklar JL, et alExpression of bcr-abl fusion transcripts following bone marrow transplantation for Philadelphia chromosome-positive leukemia. Leukemia 4: 541, 1990.PubMedGoogle Scholar
  11. 11.
    . Hughes TP, Morgan GJ, Martiat P, et alDetection of residual leukemia after bone marrow transplant for chronic myeloid leukemia: Role of polymerase chain reaction in predicting relapse. Blood 77: 874, 1991.PubMedGoogle Scholar
  12. 12.
    . Gribben JG, Freedman AS, Neuberg D, et alImmunologic purging of marrow assessed by PCR before autologous bone marrow transplantation for B-cell lymphoma. N Engl J Med 325: 1525, 1991.PubMedCrossRefGoogle Scholar
  13. 13.
    . Brisco MJ, Condon J, Hughes E, et alOutcome prediction in childhood acute lymphoblastic leukaemia by molecular quantification of residual disease at the end of induction. Lancet 343: 196, 1994.PubMedCrossRefGoogle Scholar
  14. 14.
    . LoCoco F, Diverio D, Pandolfi PP, et alMolecular evaluation of residual disease as a predictor of relapse in acute promyelocytic leukaemia. Lancet 340: 1437, 1992.PubMedCrossRefGoogle Scholar
  15. 15.
    . Huang W, Sun G-L, Li X-S, et alAcute promyelocytic leukemia: Clinical relevance of two major PML-RARα isoforms and detection of minimal residual disease by retrotranscriptase/polymerase chain reaction to predict relapse. Blood 82: 1264, 1993.PubMedGoogle Scholar
  16. 16.
    . Miller WH Jr, Levine K, DeBlasio A, et al. Detection of minimal residual disease in acute promyelocytic leukemia by a reverse transcriptase polymerase chain reaction assay for the PML/RAR-α fusion mRNA. Blood 82: 1689, 1993.PubMedGoogle Scholar
  17. 17.
    . Thompson JD, Brodsky I & Yunis JJ. Molecular quantification of residual disease in chronic myelogenous leukemia after bone marrow transplantation. Blood 79: 1629, 1992.PubMedGoogle Scholar
  18. 18.
    . Malinge MC, Mahon FX, Delfau MH, Daheron L, Kitzis A, Guilhot F, Tanzer J, Grandchamp B. Quantitative determination of the hybrid Bcr-Abl RNA in patients with chronic myelogenous leukaemia under interferon therapy. Brit J Haematol 82: 701, 1992.CrossRefGoogle Scholar
  19. 19.
    . Lion T, Henn T, Gaiger A, et alEarly detection of relapse after bone marrow transplantation in patients with chronic myelogenous leukaemia. Lancet 341: 275, 1993.PubMedCrossRefGoogle Scholar
  20. 20.
    . Cross NC, Feng L, Chase A, et alCompetitive polymerase chain reaction to estimate the number of BCR-ABL transcripts in chronic myeloid leukemia patients after bone marrow transplantation. Blood 82: 1929, 193.Google Scholar
  21. 21.
    . Lion T. Clinical implications of qualitative and quantitative polymerase chain reaction analysis in the monitoring of patients with chronic myelogenous leukemia. The European Investigators on Chronic Myeloid Leukemia Group. Bone Marrow Transplantation 14: 505, 1994.PubMedGoogle Scholar

Copyright information

© Plenum Press, New York 1996

Authors and Affiliations

  • Harry Iland
    • 1
    • 2
  • Francisca Springall
    • 1
  • Tao Zeng
    • 1
  • Katrina Bradfield
    • 1
  1. 1.The Kanematsu LaboratoriesRoyal Prince Alfred HospitalCamperdownAustralia
  2. 2.Kanematsu LaboratoriesRoyal Prince Alfred HospitalCamperdownAustralia

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