《中华血液学杂志》
2023年第11期
DOI:
10.3760/cma.j.issn.0253-2727.2023.11.004
高汉林,郝玥,陈文敏,李玲娣,王旭,秦亚溱,江倩
Abstract
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 水平并纳入统计分析。整体而言,两种方法检测结果差异具有统计学意义(P < 0.001)。按分子学反应分层分析显示,该差异仅存在于达到 MR4.5 及以上水平者(P < 0.001);未达主要分子学反应(MMR)(P = 0.922)、达到 MMR(P = 0.723)及分子学反应 4(MR4)(P = 0.099)层级均无明显差异。总体分析可见,dPCR 与 qPCR 检测 BCR::ABL mRNA 水平呈中度相关(r = 0.761,P < 0.001)。随分子学反应程度加深,二者相关性逐步降低直至无明显相关: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)。一致性分析结果提示,MR4.5 组两种检测方法的一致性显著差于其余分层:未达 MMR(r = 0.042,P = 0.846)、MMR(r = 0.054,P = 0.229)、MR4(r = −0.020,P = 0.399)、MR4.5(r = −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。
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: r = 0.042, P = 0.846; MMR: r = 0.054, P = 0.229; MR4: r = -0.020, P = 0.399; MR4.5: r = -0.219, P < 0.001) .
Conclusion:
dPCR is more accurate than qPCR for measuring BCR::ABL (P210) mRNA expression in patients with CML who achieve a stable deep MR.
Tags:
白血病,髓样,慢性(Leukemia, myeloid, chronic); 实时定量聚合酶链反应( Real- time quantitative polymerase chain reaction); 数字聚合酶链反应(Digital polymerase chain reaction); 融合蛋白质类,BCR::ABL(Fusion proteins, BCR::ABL)