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儿童急性淋巴细胞白血病微量残留病细胞的DNA定量研究 |
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卢洁 谷仁凯 佟琳如 庞秀英 孙立荣 李学荣 董增义 宋如俊
【摘要】 目的 探讨定量监测儿童急性淋巴细胞白血病（ALL）微量残留病（MRD）细胞DNA变化的临床意义。方法 采用5′端修饰引物，构建内参照竞争模板，建立竞争性聚合酶链反应（competitive polymerase chain reaction, CPCR）DNA(CPCR-DNA)定量法，对70例ALL患儿进行了T细胞受体（T-cell receptor，TCR）Vδ2Dδ3基因重排定性检测及完全缓解（CR）期MRD的动态定量研究。结果 (1)TCRVδ2Dδ3基因重排检出率：在70例ALL中占81%，48例B-ALL中占83%，4例T-ALL中占1/4，后两者比较差异有显著意义（P＜0.05）。(2)MRD-DNA定量监测显示：①随CR期延长（除CR<3个月者），MRD水平呈逐年下降趋势，以CR 4～12个月MRD水平最高，并与骨髓复发呈正相关。②9例复发者于复发前平均7个月即出现MRD增高，分子水平的改变先于临床复发。③ALL持续完全缓解（CCR）3年以上，MRD持续转阴或持续≤0.05%，可作为终止化疗的可靠指标。结论 (1)建立了一种简捷、敏感、准确的CPCR-DNA定量法;(2)用该CPCR方法定量监测MRD变化，对判断疗效、预测复发、指导治疗和决定终止化疗时间有一定的临床意义。
【关键词】 白血病，淋巴细胞，急性 肿瘤，残留 DNA 聚合酶链反应 序列分析
Quantitative analysis of minimal residual disease cells DNA in children with acute lymphoblastic leukemia LU Jie, GU Renkai, TONG Linru, et al. Department of Pediatrics, Affiliated Hospital, Medical School of Qingdao University, Qingdao 266003
【Abstract】 Objective To explore the clinical significance of quantitative analysis of minimal residual disease (MRD) cells DNA in children with acute lymphoblastic leukemia (ALL). Methods By using the competitive polymerase chain reaction (CPCR) technique which was based on the 5′-terminal modified primers to obtain the internal standard template, the T-cell receptor (TCR)Vδ2Dδ3 gene rearrangements were detected in 162 bone morrow samples collected from 70 patients with ALL, and MRD-DNA were analysed quantitatively in patients with complete remission (CR). Results (1) The frequencies of TCR Vδ2Dδ3 rearrangements were 81% in ALL, 83% in B-ALL and 1/4 in T-ALL, respectively. (2)MRD-DNA was detected in 92 samples collected from 48 patients with CR, and the MRD showed a descending tendency year by year in the patients, but not the patients with CR less than 3 months. ① The highest MRD DNA level was found in the patients with CR of 4～12 months and showed a positive correlation with the re[1] [2] 下一页
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