高通量测序在FLT3-ITD阳性急性髓细胞白血病患者中的应用

发布时间：2018-01-06 14:18

  本文关键词：高通量测序在FLT3-ITD阳性急性髓细胞白血病患者中的应用　出处：《中国实验血液学杂志》2016年02期 　论文类型：期刊论文

  更多相关文章： 高通量测序 FLT基因 急性髓细胞白血病 毛细管电泳 ITD突变

【摘要】：目的:探讨高通量测序在FLT3基因ITD突变含量较低及存在多条ITD突变的初诊急性髓细胞白血病(AML)患者ITD序列分析中的应用,并分析ITD突变特点。方法:采用聚合酶链反应(PCR)扩增23例FLT3-ITD阳性AML患者FLT3基因,应用毛细管电泳法分析FLT3基因ITD突变,然后用带有不同序列标签的PCR引物再次扩增上述患者FLT3基因,应用illumina Miseq二代测序仪分析扩增产物,并将测序结果与UCSC数据库比对。结果:23例AML患者毛细管电泳检测显示,17例患者为1条ITD插入突变,3例患者为2条ITD插入突变,3例患者为3条ITD插入突变。高通量测序共检测到的33条ITD中17条ITD插入序列为FLT3野生型完全重复,16条ITD为野生型部分重复。1例伴多条ITD患者同一插入长度存在两种ITD序列,另外1例伴有2条ITD为同一ITD插入位点。ITD插入序列发生在p.Y572位至p.L602位之间,所有患者ITD均覆盖p.V592-p.E598之间的一个或多个氨基酸。结论:Illumina Miseq二代测序仪可灵敏、准确地分析FLT3-ITD序列,FLT3-ITD序列特征变化较大但突变热点区域较为集中。
[Abstract]:Objective: To investigate the high-throughput sequencing in FLT3 gene ITD mutation was lower and the presence of multiple ITD mutations in primary acute myeloid leukemia (AML) patients with application analysis in the ITD sequence, and analyze the characteristic of ITD mutation. Methods: polymerase chain reaction (PCR) amplification in 23 cases of FLT3-ITD positive patients with AML FLT3 gene analysis. FLT3 gene ITD mutation by capillary electrophoresis, and then use the PCR primers with different sequence tags again amplified the FLT3 gene of patients, using Illumina Miseq two generation sequencing analysis of PCR products, and the sequencing results with UCSC database. Results: the results showed that AML patients were detected by capillary electrophoresis in 23 cases, 17 cases of patients with 1 ITD insertion mutations in 3 patients with 2 ITD insertions, 3 patients with 3 ITD insertion mutation. 33 ITD high-throughput sequencing were detected in 17 ITD insertion sequence FLT3 complete duplication of the wild type, 16 wild type ITD Repeat.1 cases with multiple ITD patients with the same insertion length there are two kinds of ITD sequences, the other 1 cases with 2 ITD for the same ITD.ITD insertion sites of insertion sequence between p.Y572 to p.L602, ITD in all patients were covered with one or more amino acids between p.V592-p.E598. Conclusion: Illumina Miseq the two generation sequencing instrument can be sensitive, accurate analysis of the FLT3-ITD sequence, FLT3-ITD sequence variation of large but hot spot mutations are more concentrated.

【作者单位】： 中日友好医院检验科;
【基金】：中日友好医院青年课题(QN-2013-11)
【分类号】：R440;R733.71
【正文快照】： Fms样酪氨酸激酶-3(fms-like tyrosine kinase-3,FLT3)是Ⅲ型受体酪氨酸激酶家族成员,在约20%-30%的急性髓系白血病(acute myeloid leuke-mia,AML)患者中FLT3基因中外显子14和15存在内部串联重复(FLT3 internal tandem duplication, FLT3-ITD)[1]。FLT3-ITD对AML患者的完全缓，

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