Quantitative real-time RT-PCR analysis of PML-RARalpha mRNA in acute promyelocytic leukemia: assessment of prognostic significance in adult patients from intergroup protocol 0129

Abstract

The potential prognostic value of quantitative real-time reverse transcriptionpolymerase chain reaction (RT-PCR [qrt-PCR]) measurements of PML-RAR␣ mRNA in acute promyelocytic leukemia was retrospectively assessed before treatment and at 3 posttreatment intervals in 123 patients on intergroup protocol 0129. The primary measure was the PML-RAR␣ GAPDH normalized quotient (NQ), that is, PML-RAR␣ mRNA copies divided by glyceraldehyde-3-phosphate dehydrogenase (GAPDH) mRNA copies. Only samples with more than 2.5 ؋ 10 5 copies of the housekeeping gene GAPDH mRNA (detection sensitivity exceeding 10 4 ) were considered NQ evaluable. With RNA from low-density selected cells, paired peripheral blood (PB) and bone marrow samples (n ‫؍‬ 140) had comparable NQs (P < .001). Before treatment, high NQ was associated with short-form PML-RAR␣ (P < .001), but not with white blood cell count or clinical outcome. Following treatment, NQ was lower in all-trans retinoic acidinduced complete remission (CR) than chemotherapy-induced CR (P ‫؍‬ .018) and at first test after consolidation chemotherapy (P ‫؍‬ .037). After consolidation chemotherapy, patients with NQ exceeding 10 ؊5 had 4.1-fold increased relapse risk (P ‫؍‬ .008); however, 73% of patients who experienced relapse had NQ lower than 10 ؊5 . In the follow-up period (FUP), any NQ exceeding 10 ؊5 and 10 ؊6 had 17.5-fold and 7.6-fold increased relapse risk, respectively (P < .001), while no gradation of relapse risk (approximately 18%) could be identified at NQ lower than 10 ؊6 , including NQ ؊ .

Quantitative Assessment of PML-RARa and BCR-ABL by Two Real-Time PCR Instruments: Multiinstitutional Laboratory Trial

“…In this study, similar to other published results obtained by gradient gel electrophoresis (13 ), the HDL particle diameter (nm) was greater in premenopausal women than in men (9.1 vs 8.4 nm; Mann-Whitney test, P ϭ 0.013). Negative correlations between HDL particle size and total cholesterol and LDL-cholesterol were also reported by Pascot et al (13 ), thus strengthening the link between HDL particle size and coronary artery disease. In the present study, the correlations between HDL particle size and HDL-cholesterol and triglyceride concentrations were not statistically significant, probably because of the small number of samples.…”

“…In the present study, the correlations between HDL particle size and HDL-cholesterol and triglyceride concentrations were not statistically significant, probably because of the small number of samples. Pascot et al (13 ) may have found significant correlations for these relationships because they studied Ͼ400 individuals. Therefore, LLS analysis after chemical precipitation of apo B-containing lipoproteins gave results for HDL diameter in the same range as those obtained by established techniques for lipoprotein sizing (9,10,13 ).…”

“…In healthy normolipidemic individuals, HDL sizing by nuclear magnetic resonance spectroscopy yielded diameters of ϳ9.2 nm (10 ), whereas the values obtained from gradient gel electrophoresis ranged from 8.4 to 9.6 nm (9,13 ). In this study, similar to other published results obtained by gradient gel electrophoresis (13 ), the HDL particle diameter (nm) was greater in premenopausal women than in men (9.1 vs 8.4 nm; Mann-Whitney test, P ϭ 0.013).…”

How to manage acute promyelocytic leukemia

“…The first sign of recurrence is usually indicated by the PCR detection of PML --RARa transcripts in bone marrow. The vast majority of patients relapse in the first 3 years after the end of treatment; 84 however, there are also individuals who only relapse in the extramedullary sites (such as the CNS).…”

Combined megaplex TCR isolation and SMART-based real-time quantitation methods for quantitating antigen-specific T cell clones in mycobacterial infection

“…To standardize expression levels of Vβ and Cβ transcripts, C T Vβ and C T Cβ were verified by C T values for a house-keeping gene, β actin (C T β actin). The expression levels of individual clonotypic TCR transcripts among a total of TCR β transcripts within the sample were then calculated using the formula, 2 −[(CTVβ −CTβactin) −(CTCβ−CTβactin)] , as previously described (Livak and Schmittgen, 2001;Semighini et al, 2002;Gallagher et al, 2003). The calculation gave rise to the ratio of clonotypic Vβ-bearing transcripts versus Cβ transcripts.…”