数字PCR与实时荧光定量PCR检测慢性髓性白血病患者BCR::ABL mRNA水平的比较性研究  

Comparison of BCR::ABL(P210)mRNA levels detected by dPCR and qPCR methods in patients with chronic myeloid leukemia

作　　者：高汉林 郝玥 陈文敏 李玲娣[1] 王旭[1] 秦亚溱[1] 江倩[1] Gao Hanlin;Hao Yue;Chen Wenmin;Li Lingdi;Wang Xu;Qin Yazhen;Jiang Qian(Peking University People's Hospital,Peking University Institute of Hematology,Beijing 100044,China)

机构地区：[1]北京大学人民医院、北京大学血液病研究所,北京100044

出　　处：《中华血液学杂志》2023年第11期906-910,共5页Chinese Journal of Hematology

基　　金：国家自然科学基金(81970140)。

摘　　要：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(d^(-)=0.042,P=0.846),MMR(d^(-)=0.054,P=0.229),MR4(d^(-)=-0.020,P=0.399),MR4.5(d^(-)=-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:d^(-)=0.042,P=0.846;MMR:d^(-)=0.054,P=0.229;MR4:d^(-)=-0.020,P=0.399;MR4.5:d^(-)=-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.

关 键 词：白血病 髓样 慢性 实时定量聚合酶链反应 数字聚合酶链反应 融合蛋白质类 BCR::ABL 

分 类 号：R733.72[医药卫生—肿瘤]