Acute promyelocytic leukemia (APL) is a medical emergency which requires rapid diagnosis and tailored treatment. Detection of the PML/RARalpha fusion gene in APL blasts is critical to start promptly the specific therapy with all-trans retinoic acid (ATRA). APL lacking this genetic lesion have been reported as being ATRA resistant. Reverse-transcription polymerase chain reaction (RT-PCR) has been extensively used to detect the PML/RARalpha cDNA. The reported PML/RARalpha amplification techniques are laborious and time consuming, and include conventional RNA extraction, cDNA synthesis and a two-round (nested) PCR. We hereby describe a few variations of the commonly adopted RNA extraction and PML/RARalpha RT-PCR protocols which allow a molecular diagnosis of APL to be carried out in less than 5 h. Processing of small volumes of leukemic cell lysate (0.5 ml) in a microfuge allows extraction of good quality RNA in 1 h. After reverse transcription to obtain cDNA, a 'hot start' PCR procedure was adopted which enabled us to amplify clearly visible and specific products after a single (not nested) amplification round. The PML/RARalpha fusion gene was detected in the blasts of six consecutive APL at diagnosis, and an APL-tailored protocol including ATRA was started in each case within 6 h of admission. On repeated experiments, the assay proved highly specific and sensitive for the rapid detection of all PML/RARalpha transcript types. Our data should encourage the use of this rapid procedure for the diagnosis of both typical APL and, particularly, less typical cases awaiting urgent therapeutic intervention.
Diverio, D., Riccioni, R., Pistilli, A., Buffolino, S., Avvisati, G., Mandelli, F., et al. (1996). Improved rapid detection of the PML/RARalpha fusion gene in acute promyelocytic leukemia. LEUKEMIA, 10(7), 1214-1216.
|Tipologia:||Articolo su rivista|
|Citazione:||Diverio, D., Riccioni, R., Pistilli, A., Buffolino, S., Avvisati, G., Mandelli, F., et al. (1996). Improved rapid detection of the PML/RARalpha fusion gene in acute promyelocytic leukemia. LEUKEMIA, 10(7), 1214-1216.|
|Settore Scientifico Disciplinare:||Settore MED/15 - Malattie del Sangue|
|Revisione (peer review):||Esperti anonimi|
|Stato di pubblicazione:||Pubblicato|
|Data di pubblicazione:||lug-1996|
|Titolo:||Improved rapid detection of the PML/RARalpha fusion gene in acute promyelocytic leukemia|
|Autori:||Diverio, D; Riccioni, R; Pistilli, A; Buffolino, S; Avvisati, G; Mandelli, F; Lo Coco, F|
|Appare nelle tipologie:||01 - Articolo su rivista|