Preliminary experience in external quality control of RT-PCR PML-RARα detection in promyelocytic leukemia

Abstract

To standardize the results obtained in PML/RAR␣ RT-PCR detection by laboratories of hospitals involved in the SpanishProgram for Treatment of Hematological Malignancies (PETHEMA) LPA-96, designed for the treatment of acute promyelocytic leukemia (APL), cDNA samples obtained by reverse transcription of RNA from bone marrow samples of patients with APL were sent to participating laboratories. During the first year of this external quality assessment trial nine samples were tested by a maximum of 12 laboratories. The control gene was satisfactorily amplified in 90% of the samples (62 of 69 samples), supporting the adequacy of the cDNA to be used as control sample. There was an 83% concordance between laboratories for PML/RAR␣ detection with similar results for the type of PML/RR␣ rearrangements. However, 17% disagreement still remained, attributable to low sensitivity or inadequacy of methods followed. The results stressed the need for implementation of an external quality assessment scheme to ensure the standardization of the results.

Quantitation of minimal residual disease in acute promyelocytic leukemia patients with t(15;17) translocation using real-time RT-PCR

“…This level of detection is a little higher than the 10 3 Leukemia copies previously reported, 7 but further confirms that PML-RAR␣ is weakly expressed as previously described. 22,23 This quantitative method will in fact be a useful tool to optimize the upstream steps of the RT-PCR reaction such as patient cell handling, anticoagulant used, shipment, RNA extraction, and reverse transcription, in order to increase the yield in cDNA, an important limiting factor specifically noted in APL cells.…”

Prognostic value of FLT3 mutations in patients with acute promyelocytic leukemia treated with all-trans retinoic acid and anthracycline monochemotherapy

“…Molecular relapse was defined as the reappearance of PCR positivity in two consecutive bone marrow samples at any time after consolidation therapy. 28 A genetic diagnosis of APL using PCR, anti-PML staining or cytogenetic tests was required for the diagnosis of overt hematologic relapse.…”

Risk-adapted treatment of acute promyelocytic leukemia with all-trans-retinoic acid and anthracycline monochemotherapy: a multicenter study by the PETHEMA group

“…RT-PCR tests were carried out by 14 different laboratories involved in an external quality control program, which included interlaboratory exchange of samples, as reported elsewhere. 12,13 If a positive PCR result was reported after consolidation or beyond, a new sample was obtained and sent to one of 2 reference laboratories (P.B. and M.G.…”