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目的微量残留病(MRD)监测是儿童急性淋巴细胞白血病(ALL)早期治疗反应中最重要的预后因素之一。目前常用的MRD检测的方法主要有流式细胞术和PCR技术,但单用一种方法均不能为所有的患儿找到合适的监测标记,该研究探讨两种方法联合应用是否能为大多数ALL患儿找到合适的MRD监测标记。方法联合应用流式细胞术和PCR技术筛选上海儿童医学中心2001年9月至2003年10月126例新发ALL患儿骨髓标本的异常免疫表型及抗原受体基因重排。结果①106例B系-ALL患儿骨髓标本用四色流式细胞术进行了MRD免疫表型标记的筛选,其中11例标本未筛选出监测标记,阳性率为89.6%;有1个监测标记的标本为11例(11.6%),至少有两个标记的标本占88.4%。②PCR技术筛选27例ALL骨髓标本抗原受体基因重排,26例至少有一个标记(占96.3%),其中9例(34.6%)骨髓标本只有一个监测标记,17例(65.4%)至少有两个监测标记。在T系-ALL骨髓标本中,以TCRVγⅠ-Jγ1.3/2.3阳性最多,双等位基因重排发生阳性率较高;系性交叉抗原表达在B系-ALL骨髓标本中表达较高,达57.1%(4/7)。③两种方法联用能为121例(96.0%)的患儿提供合适的筛选指标。结论流式细胞技术检测异常免疫表型与PCR技术检测抗原受体基因重排联用,可为绝大多数的ALL患儿找到合适的MRD监测指标;在抗原受体基因重排中,存在系性交叉抗原表达及双等位基因重排。
Objective Microdisorders (MRD) monitoring is one of the most important prognostic factors in the early response to childhood acute lymphoblastic leukemia (ALL). Currently used MRD detection methods are mainly flow cytometry and PCR technology, but only one method can not find a suitable monitoring marker for all children, the study to explore whether the combination of two methods for most ALL Children found the right MRD monitoring mark. Methods The abnormal immunophenotypes and antigen receptor rearrangements of 126 newly diagnosed ALL children with bone marrow from Shanghai Children’s Medical Center from September 2001 to October 2003 were screened by flow cytometry and PCR. Results ① The bone marrow samples from 106 children with B-ALL were screened by four-color flow cytometry with the immunophenotypic markers of MRD. Among them, 11 samples were not screened, the positive rate was 89.6% The specimens were 11 (11.6%), at least two labeled specimens accounted for 88.4%. PCR was used to screen the gene rearrangement of antigen receptor in 27 cases of ALL bone marrow samples. There were at least one marker (96.3%) in 26 cases, of which 9 cases (34.6%) had only one monitoring marker and 17 cases (65.4%) had at least two markers A monitoring mark. In T-ALL bone marrow samples, the positive rate of TCRVγⅠ-Jγ1.3 / 2.3 was the highest, and the positive rate of biallelic rearrangement was higher. The expression of tethered antigen was higher in B line-ALL bone marrow 57.1% (4/7). ③ The two methods combined with 121 cases (96.0%) for children with appropriate screening indicators. Conclusion The detection of abnormal immunophenotype by flow cytometry and PCR detection of antigen receptor gene rearrangement can find the suitable MRD monitoring index for the majority of children with ALL. In the gene rearrangement of antigen receptor, Sexual cross antigen expression and double allelic rearrangement.