关键词: Digital polymerase chain reaction Fusion proteins, BCR::ABL Leukemia, myeloid, chronic Real-time quantitative polymerase chain reaction

Mesh : Humans Cross-Sectional Studies Cytogenetics Leukemia, Myelogenous, Chronic, BCR-ABL Positive / genetics Real-Time Polymerase Chain Reaction RNA, Messenger / genetics

来  源:   DOI:10.3760/cma.j.issn.0253-2727.2023.11.004   PDF(Pubmed)

Abstract:
Objective: To compare digital polymerase chain reaction (dPCR) and real-time quantitative PCR (qPCR) measurements of BCR::ABL (P210) mRNA expression in patients with chronic myeloid leukemia (CML) . Methods: In this non-interventional, cross-sectional study, BCR::ABL (P210) mRNA was simultaneously measured by dPCR and qPCR in peripheral blood samples collected from patients with CML who underwent tyrosine kinase inhibitor therapy and who achieved at least a complete cytogenetic response from September 2021 to February 2023 at Peking University People\'s Hospital. The difference, correlation, and agreement between the two methods were evaluated using the Wilcoxon signed-rank test, Spearman\'s correlation, and Bland-Altman analysis, respectively. Results: In total, 459 data pairs for BCR::ABL mRNA expression measured by dPCR and qPCR from 356 patients with CML were analyzed. There was a significant difference in BCR::ABL mRNA expression between the two methods (P<0.001). When analyzed by the depth of the molecular response (MR), a significant difference only existed for patients with ≥MR4.5 (P<0.001). No significant difference was observed for those who did not achieve a major MR (no MMR; P=0.922) or for those who achieved a major MR (MMR; P=0.723) or MR4 (P=0.099). There was a moderate correlation between the BCR::ABL mRNA expression between the two methods (r=0.761, P<0.001). However, the correlation gradually weakened or disappeared as the depth of the MR increased (no MMR: r=0.929, P<0.001; MMR: r=0.815, P<0.001; MR4: r=0.408, P<0.001; MR4.5: r=0.176, P=0.176). In addition, the agreement in BCR::ABL mRNA expression between the two methods in those with MR4.5 was weaker than other groups (no MMR: ▉= 0.042, P=0.846; MMR:▉=0.054, P=0.229; MR4:▉=-0.020, P=0.399; MR4.5:▉=-0.219, P<0.001) . Conclusions: dPCR is more accurate than qPCR for measuring BCR::ABL (P210) mRNA expression in patients with CML who achieve a stable deep MR.
目的: 比较数字PCR(dPCR)与实时荧光定量PCR(qPCR)方法检测慢性髓性白血病(CML)患者外周血BCR::ABL(P210) mRNA水平的差异性、相关性与一致性。 方法: 本研究为非干预性、横断面研究。收集2021年9月至2023年2月在北京大学人民医院就诊的服用酪氨酸激酶抑制剂(TKI)至少获得完全细胞遗传学反应(CCyR)的CML患者同时采用dPCR和qPCR方法检测BCR::ABL mRNA水平的结果,分别采用Wilcoxon符号秩检验、Spearman相关系数、Bland-Altman分析比较两种方法的差异性、相关性、一致性。 结果: 356例CML患者的459对外周血样本分别采用dPCR与qPCR方法检测BCR::ABL mRNA水平的结果用于分析。总体上,两种方法检测结果差异有统计学意义(P<0.001)。根据分子学反应分层分析,这种差异仅存在于获得≥分子学反应4.5(MR4.5)(P<0.001)后,而非未达主要分子学反应(MMR)(P=0.922)、MMR(P=0.723)和分子学反应4(MR4)(P=0.099)水平。总体上,dPCR与qPCR方法检测BCR::ABL mRNA水平具有中度相关性(r=0.761,P<0.001)。但随着分子学反应加深,这种相关性逐渐减弱甚至消失:未达MMR(r=0.929,P<0.001),MMR(r=0.815,P<0.001),MR4(r=0.408,P<0.001),MR4.5(r=0.176,P=0.176)。一致性上,MR4.5组两种方法检测结果的一致性弱于其他各组:未达MMR(▉=0.042,P=0.846),MMR(▉=0.054,P=0.229),MR4(▉=-0.020,P=0.399),MR4.5(▉=-0.219,P<0.001)。 结论: CML患者当获得深层分子学反应后,更适合采用dPCR方法监测BCR::ABL(P210) mRNA水平以提高精准性。.
摘要:
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