|Detecting PML-RAR alpha transcript in acute promyelocytic leukemia using real-time quantitative RT-PCR|
|Zhu Hong-Hu; Liu Yan-Rong; Qin Ya-Zhen; Bin, Jiang; Shan Fu-Xiang; Wu Shu-Lan; Yang Ping-Di; Re, Zhao; Lu Dao-Pei|
|关键词||real-time quantitative RT-PCR arsenics all-trans retinoic acid acute promyelocytic leukemia PML-RAR alpha|
|刊名||CHINESE MEDICAL JOURNAL|
|WOS标题词||Science & Technology|
|类目[WOS]||Medicine, General & Internal|
|研究领域[WOS]||General & Internal Medicine|
|关键词[WOS]||POLYMERASE-CHAIN-REACTION ; RESIDUAL DISEASE DETECTION ; CANCER PROGRAM ; RETINOIC ACID ; FUSION GENE ; DIAGNOSIS ; MULTICENTER ; MANAGEMENT ; REMISSION ; SURVIVAL|
Background Real-time quantitative RT-PCR (RQ-PCR) assay has become a vital tool to monitor residual disease of leukemia. However, the complexity and standardization of RQ-PCR should never be overlooked and the results should be interpreted cautiously in clinical conditions. We aimed to assess the methodology of RQ-PCR and its clinical applications in monitoring molecular kinetics of 36 newly diagnosed cases of acute promyelocytic leukemia patients with t (15; 17) from October 2004 to December 2005.
Methods All the TaqMan probe-based RQ-PCR reactions and analysis were performed on an ABI-PRISM 7500 platform. The quantitation of PML-RAR alpha transcripts was represented by the normalized quotient, that is, PML-RAR alpha transcript copies divided by ABL transcript copies. According to induction therapy, the patients were classed into two groups: group 1 (n=23), three-drug combination including arsenics, all-trans retinoic acid and mitoxantrone; and group 2 (n=13), two-drug combination from all-trans retinoic acid, arsenics and mitoxantrone.
Results The sensitivity of RQ-PCR was 1 per 105 Cells and 5 copies of the PML-RAR alpha transcript could be reproducibly detected. No false positive results occurred in 40 non-acute promyelocytic leukemia samples. Optimal amplification efficiency could be attained, which was determined by the slope of the standard curves (slope: -3.2 - -3.7). The inter-assay and intra-assay variation coefficients of the method were 1.01 % and 0.56% respectively. Although the time to attain hematological complete remission was similar in both groups, the time to achieve molecular remission of group 1 was significantly shorter than that of group 2 (61 days vs; 75 days, P=0.034). The rate of molecular remission within 70 days was higher in group 1 than in group 2 (75.00% vs; 38.46%, P=0.036). Compared with pretreatment, median reduction of the PMIL-RAR alpha transcript before first consolidation therapy differed significantly between group 1 and group 2 (log scale, 3.15 vs 2.31, P=0.024). Interestingly, we found that PML-RAR alpha transcript levels temporarily increased in bone marrow (7 patients) and peripheral blood (22 patients) samples of patients during induction therapy in both groups.
Conclusions The RQ-PCR assay is reliable for the detection of PML-RAR alpha transcripts. Arsenics, all-trans retinoic acid and mitoxantrone triad induction treatment of acute promyelocytic leukemia is superior to two-drug combination induction therapy in terms of the molecular response.
|作者单位||1.Peking Univ, Peoples Hosp, Inst Hematol, Beijing 100044, Peoples R China|
2.Beijing Daopei Hosp, Beijing 100049, Peoples R China
|Zhu Hong-Hu,Liu Yan-Rong,Qin Ya-Zhen,et al. Detecting PML-RAR alpha transcript in acute promyelocytic leukemia using real-time quantitative RT-PCR[J]. CHINESE MEDICAL JOURNAL,2007,120(20):1803-1808.|
|APA||Zhu Hong-Hu.,Liu Yan-Rong.,Qin Ya-Zhen.,Bin, Jiang.,Shan Fu-Xiang.,...&Lu Dao-Pei.(2007).Detecting PML-RAR alpha transcript in acute promyelocytic leukemia using real-time quantitative RT-PCR.CHINESE MEDICAL JOURNAL,120(20),1803-1808.|
|MLA||Zhu Hong-Hu,et al."Detecting PML-RAR alpha transcript in acute promyelocytic leukemia using real-time quantitative RT-PCR".CHINESE MEDICAL JOURNAL 120.20(2007):1803-1808.|