论文部分内容阅读
目的探讨运用自动片段分析的多重PCR检测IgH和(或)TCRγ基因单克隆重排作为检测儿童急性淋巴细胞白血病(ALL)微小残留病(MRD)方法的可行性。方法采用1∶3匹配设计的病例-对照研究方法,收集2009年1月至2011年12月中山大学附属第一医院4例极早期复发的非高危儿童ALL(病例组)和危险度匹配的12例非高危儿童ALL(对照组)在初诊时、诱导化疗结束(距开始治疗5周,W5)、再诱导化疗前(距开始治疗23周,W23)及维持第3个月(距开始治疗44周,W44)的骨髓样本,治疗方案均按照广东2008ALL协作组方案进行。用多重PCR方法定性检测各时间两组骨髓样本是否存在IgH和(或)TCRγ基因单克隆重排,PCR产物采用片段分析软件进行分析。检出单克隆重排的IgH和(或)TCRγ被认为是MRD阳性。结果复发组4例和对照组12例ALL患儿在初诊时均监测到IgH单克隆重排,TCRγ单克隆重排阳性率56%。在W5时,病例组和对照组的MRD阳性率的差异无统计学意义(P<0.05)。在W23和W44这两个时间点,病例组的MRD阳性率显著高于对照组,差异有统计学意义(P<0.05)。结论多重PCR联合自动片段分析检测IgH和(或)TCRγ基因单克隆重排的方法可以用于儿童ALL治疗过程中的MRD监测。
Objective To investigate the feasibility of using multiplex PCR to detect the monoclonal rearrangement of IgH and / or TCRγ gene as a method of detecting minimal residual disease (MRD) in children with acute lymphoblastic leukemia (ALL). Methods A 1: 3 matched case-control study was conducted in 4 non-high risk childhood ALL patients (case group) and 12 matched hazard patients from January 2009 to December 2011 in the First Affiliated Hospital of Sun Yat-sen University Cases of non-high-risk children ALL (control group) at the time of the first induction chemotherapy induction (5 weeks from the beginning of treatment, W5), before induction of chemotherapy (23 weeks after the start of treatment, W23) and maintain the first 3 months Week, W44) bone marrow samples, treatment programs are in accordance with Guangdong 2008ALL collaborative group program. Multiplex PCR was used to qualitatively detect the presence of IgH and / or TCRγ gene monoclonal rearrangements in two groups of bone marrow samples at each time point. PCR products were analyzed by fragment analysis software. IgH and / or TCRγ, which detected a monoclonal rearrangement, were considered MRD positive. Results The IgH monoclonal rearrangement was detected in 4 cases of relapsed group and 12 cases of ALL in control group. The positive rate of TCRγ monoclonal rearrangement was 56%. At W5, there was no significant difference in the MRD positive rate between the case group and the control group (P <0.05). At the two time points W23 and W44, the positive rate of MRD in case group was significantly higher than that in control group (P <0.05). Conclusion The multiplex PCR combined with auto-segment analysis to detect IgH and / or TCRγ gene monoclonal rearrangement can be used to monitor MRD in childhood ALL.