CAJ | 학술논문

目的探索自动化片段分析的多重聚合酶链反应(PCR)检测免疫球蛋白重链(IgH)和T淋巴细胞受体γ(TCRγ)用于监测儿童急性淋巴细胞白血病(ALL)微小残留病(MRD)的可行性.方法收集自2009年5月至2013年8月在中山大学附属第一医院儿科诊治的86例ALL患儿治疗前骨髓,用定性的多重PCR方法扩增IgH和TCRγ,PCR产物采用片段分析软件GENEMAPPERID进行自动化分析.检出单克隆IgH和/或TCRγ重排的患儿,诱导结束时(D33)再次检测到单克隆重排者定为MRD阳性.所有患儿接受GD2008协作组方案治疗,随访时间截至2014年5月31日.用Kaplan-Merier法预测无事件生存率(EFS)和生存曲线,生存曲线差别用log-rank检验.结果 94.2％的患儿(81/86例)至少检测到1种阳性的IgH/TCRγ,实际进行后续MRD监测的有79例,中位随访时间20个月(9～61个月).在79例ALL患儿中,MRD阳性20例,MRD阴性59例,3年EFS分别为56.4％±14.7％和94.0％±3.4％(x2=8.563,P=0.003).根据传统的预后分型,79例MRD监测组患儿中65例为非高危患儿:标危组23例,中危组42例,3年EFS分别为95.3％±4.7％和76.6％±9.O％(x2 =0.934,P=0.334);此65例非高危患儿如果用MRD进行分层,MRD阴性组52例,MRD阳性组13例,3年EFS分别为93.1％±3.8％和59.5％±16.2％(x2=7.128,P=0.008).结论自动化片段分析的多重PCR检测IgH/TCRγ是简便有效的儿童ALL的MRD监测方法.诱导结束时MRD的监测比传统预后分型能更精确判断预后,值得积累更多的病例以得出更可靠的结论.
목적 탐색자동화편단분석적다중취합매련반응(PCR)검측면역구단백중련(IgH)화T림파세포수체γ(TCRγ)용우감측인동급성림파세포백혈병(ALL)미소잔류병(MRD)적가행성.방법 수집자2009년5월지2013년8월재중산대학부속제일의원인과진치적86례ALL환인치료전골수,용정성적다중PCR방법확증IgH화TCRγ,PCR산물채용편단분석연건GENEMAPPERID진행자동화분석.검출단극륭IgH화/혹TCRγ중배적환인,유도결속시(D33)재차검측도단극륭중배자정위MRD양성.소유환인접수GD2008협작조방안치료,수방시간절지2014년5월31일.용Kaplan-Merier법예측무사건생존솔(EFS)화생존곡선,생존곡선차별용log-rank검험.결과 94.2％적환인(81/86례)지소검측도1충양성적IgH/TCRγ,실제진행후속MRD감측적유79례,중위수방시간20개월(9～61개월).재79례ALL환인중,MRD양성20례,MRD음성59례,3년EFS분별위56.4％±14.7％화94.0％±3.4％(x2=8.563,P=0.003).근거전통적예후분형,79례MRD감측조환인중65례위비고위환인:표위조23례,중위조42례,3년EFS분별위95.3％±4.7％화76.6％±9.O％(x2 =0.934,P=0.334);차65례비고위환인여과용MRD진행분층,MRD음성조52례,MRD양성조13례,3년EFS분별위93.1％±3.8％화59.5％±16.2％(x2=7.128,P=0.008).결론 자동화편단분석적다중PCR검측IgH/TCRγ시간편유효적인동ALL적MRD감측방법.유도결속시MRD적감측비전통예후분형능경정학판단예후,치득적루경다적병례이득출경가고적결론.
Objective To explore the prognostic significance in monitoring minimal residual disease (MRD) in childhood acute lymphoblastic leukemia (ALL) by a simple method,and to detect cloned immunoglobulin H (IgH) and T cell receptor γ (TCRγ) gene rearrangements by using multiplex polymerase chain reaction (PCR) and automated fragment analysis.Methods Bone marrow samples were collected from 86 newly diagnosed cases of childhood ALL at the Department of Pediatrics,the First Affiliated Hospital of Sun Yat-Sen University,from May of 2009 to August of 2013.IgH and TCRγ gene rearrangements were amplified by qualitative multiplex PCR.The clonality of PCR production was analyzed by GENEMAPPERID software.Only those carried monoclonal IgH/TCRγ on diagnosis were arranged to monitor MRD.Detectable monoclonal IgH/TCRγby the end of induction was defined as MRD positive.All patients were treated with GD2008 ALL protocol.Clinical data of all newly-diagnosed ALL patients in the corresponding period were reviewed.The final follow-up on May 31,2014.Survival rates and event free survival (EFS) curves were estimated by the Kaplan-Meier,and compared by using the log-rank test.Results The percent age of 94.2 (81/86 cases) patients was at least 1 marker positive.Subsequent MRD was monitored in 79 cases.The median follow-up time was 20 months (9-61 months).By the end of induction,20 cases were MRD positive and 59 cases were M RD negative,and the 3-year EFS were 56.4％ ± 14.7％ and 94.0％ ± 3.4％ (x2 =8.563,P =0.003),respectively.According to the traditional prognostic stratification criteria,MRD was detected 65 cases in the non-high risk group:23 cases in standard risk group and 42 cases in intermediate risk group,and the difference of 3-year EFS had no statistical significance (95.3％ ±4.7％ vs 76.6％ ±9.0％,x2 =0.934,P =0.334).While using MRD by the end of induction as a risk stratification criterion,there was a statistical significant difference compared with the 3-year EFS for MRD-negative (n =52) group and MRD-positive (n =13) group (93.1％ ± 3.8％ and 59.5％ ± 16.2％,x2 =7.128,P =0.008).Conclusions It is a simple but feasible method to monitor MRD in childhood ALL by using this qualitative multiplex PCR with automated fragment analysis for monoclonal IgH/TCRγ gene rearrangements.MRD by the end of induction can be used as a more accurate risk stratification criterion than the traditional one.It is worth of further research.