Quantitative RT-PCR of PML-RARα Transcripts in Acute Promyelocytic Leukemia
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.
KeywordsChronic Myeloid Leukemia Acute Promyelocytic Leukemia Minimal Residual Disease Transferrin Receptor Fusion Transcript
Unable to display preview. Download preview PDF.
- 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
- 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
- 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