儿童急性B淋巴细胞白血病TEL-AML1和BCR-ABL融合基因检测与应用

发布时间：2018-07-03 07:34

本文选题：儿童急性淋巴细胞白血病 + 染色体核型分析　；参考：《南昌大学》2012年硕士论文

【摘要】：目的：分析儿童急性B淋巴细胞白血病(B-lineage acute lymphoblastic leukemia,B-ALL)TEL-AML1和BCR-ABL融合基因表达水平与患者的临床特征、细胞遗传学特征以及预后的关联情况。方法：收集2011年1月至2012年3月江西省儿童医院血液科收治的93例初诊儿童B-ALL患者骨髓标本,进行骨髓染色体G显带核型分析、流式细胞术免疫分型，，应用巢式PCR技术检测TEL-AML1和BCR-ABL融合基因，对检测结果阳性的标本采用荧光实时定量PCR进行融合基因定量检测；并检测首次诱导治疗末（d33）融合基因表达量。结果： 1、发病情况及临床特征：运用巢式PCR技术检测93例初诊B-ALL患儿骨髓标本，发现17例TEL-AML1阳性，9例BCR-ABL阳性，阳性率分别为18.28%和9.68%。根据检测结果将患者分为TEL-AML1阳性组、BCR-ABL阳性组和阴性组，三组间性别构成比、发病年龄、诱导治疗期末CR率均无统计学差异。三组间白血病细胞髓外浸润发生率、初诊外周血白细胞计数及骨髓原幼细胞占有核细胞比例存在统计学差异。 2、染色体G显带核型分析结果示：TEL-AML1阳性组患者均未检测到t(12;21)染色体易位，9例BCR-ABL融合基因阳性患者中有6例检测到t(9;22)染色体易位；阴性组中检测到2例t(8;14)染色体易位，其余未见染色体形态异常；三组间染色体超二倍体及亚二倍体比例无统计学差异，超二倍体以及亚二倍体患者在d33CR情况无统计学差异。 3、流式细胞术免疫分型结果示：三组患者CommonB-ALL比例分别为76.5%、44.4%、67.2%；PreB-ALL比例分别为23.5%、33.3%、25.4%；ProB-ALL比例分别为0、22.2%、7.5%；三组患者免疫分型构成比无统计学差异，TEL-AML1阳性组和BCR-ABL阳性组髓系抗原表达率高于阴性组。 4、RQ-PCR检测结果示： TEL-AML1阳性组融合基因表达水平范围2.01%~454.89%，BCR-ABL阳性组融合基因表达水平范围2.94%~1208.62%；BCR-ABL融合基因表达水平高于TEL-AML1融合基因表达水平。在首次诱导治疗末期（d33）TEL-AML1融合基因阳性组获得14例骨髓标本中有5例MRD阳性；BCR-ABL阳性组7例标本中有3例MRD阳性。结论: 1、本研究应用多重巢式RT-PCR检测93例儿童B-ALL患者初诊骨髓标本，检测到TEL-AML1阳性14例和BCR-ABL融合基因阳性9例，阳性率分别为18.28%和9.68%；RQ-PCR的阳性检出率与巢式PCR检测结果一致，具有良好的临床适用性； 2、TEL-AML1阳性患者与BCR-ABL阳性患者在短期预后上无明显差异，但BCR-ABL阳性组初诊时白血病细胞肿瘤负荷高于TEL-AML1阳性组和融合基因阴性组；
[Abstract]:Objective: to investigate the relationship between the expression of TEL-AML1 and BCR-ABL fusion genes in children with B-lineage acute lymphoblastic leukemia (B-ALL) and their clinical, cytogenetic and prognostic characteristics. Methods: from January 2011 to March 2012, the bone marrow samples of 93 newly diagnosed children with B-ALL were collected from Department of Hematology, Jiangxi Children's Hospital, and the karyotype of bone marrow chromosome G-banding was analyzed. The fusion genes of TEL-AML1 and BCR-ABL were detected by nested polymerase chain reaction (NPCR), the fusion genes were quantitatively detected by fluorescence real-time PCR and the expression of fusion genes was detected at the end of the first induction therapy (d33). Results: 1. The incidence and clinical characteristics: bone marrow specimens of 93 children with newly diagnosed B-ALL were detected by nested PCR. 17 cases (18.28%) were found to be positive for TEL-AML1 and 9 cases (9.68%) were positive for BCR-ABL. The patients were divided into BCR-ABL positive group and negative group according to the test results. There was no statistical difference among the three groups in sex composition, age of onset and CR rate at the end of induction therapy. The incidence of extramedullary infiltration of leukemic cells in the three groups, There were significant differences in the number of peripheral white blood cells and the percentage of nuclear cells occupied by bone marrow promyelocytes. 2. The karyotype analysis of chromosome G banding showed that no t (1221) chromosomal translocation was detected in 9 cases in the positive group of 10% TEL-AML1. In 6 patients with BCR-ABL fusion gene positive, t _ (9N _ 2) chromosome translocation was detected. In the negative group, 2 cases of t (8P14) chromosome translocation were detected, while the other cases were not abnormal in shape, and the proportion of hyperdiploid and subdiploid was not significantly different among the three groups. There was no statistical difference between hyperdiploid and subdiploid patients on d33CR. 3The results of flow cytometry showed that: the ratio of common B-ALL in the three groups was 76.50.The ratio of common B-ALL was 76.50.45%. The ratio of PreB-ALL to ProB-ALL was 23.533.33.35% ProB-ALL was 0.22.2mg / L and 7.5g / L, respectively. The expression rate of myeloid antigen in TEL-AML1 positive group and BCR-ABL positive group was higher than that in BCR-ABL positive group. 4 the results of RQ-PCR showed that the expression level of fusion gene in TEL-AML1 positive group was 2.01and 454.89% in BCR-ABL positive group. The expression level of synthase gene was 2.94 and 1208.62; The expression level of BCR-ABL fusion gene was higher than that of TEL-AML1 fusion gene. At the end of the first induction therapy (d33) TEL-AML1 fusion gene positive group, 5 of 14 bone marrow specimens were MRD positive and 3 of 7 cases were MRD positive. Conclusion: 1. In this study, the bone marrow specimens of 93 children with B-ALL were detected by multiplex nested RT-PCR. The positive rates of TEL-AML1 and BCR-ABL fusion gene were 18.28% and 9.68%, respectively. The positive rate of RQ-PCR was consistent with that of nested PCR, and had good clinical applicability, and there was no significant difference in short-term prognosis between patients with positive RQ-PCR and those with positive BCR-ABL. However, the tumor load of leukemic cells in BCR-ABL positive group was higher than that in TEL-AML1 positive group and fusion gene negative group.
【学位授予单位】：南昌大学
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R733.7

【共引文献】