本文關(guān)鍵詞：高通量測(cè)序在FLT3-ITD陽(yáng)性急性髓細(xì)胞白血病患者中的應(yīng)用　出處：《中國(guó)實(shí)驗(yàn)血液學(xué)雜志》2016年02期 　論文類型：期刊論文

  更多相關(guān)文章： 高通量測(cè)序 FLT基因 急性髓細(xì)胞白血病 毛細(xì)管電泳 ITD突變

【摘要】：目的:探討高通量測(cè)序在FLT3基因ITD突變含量較低及存在多條ITD突變的初診急性髓細(xì)胞白血病(AML)患者ITD序列分析中的應(yīng)用,并分析ITD突變特點(diǎn)。方法:采用聚合酶鏈反應(yīng)(PCR)擴(kuò)增23例FLT3-ITD陽(yáng)性AML患者FLT3基因,應(yīng)用毛細(xì)管電泳法分析FLT3基因ITD突變,然后用帶有不同序列標(biāo)簽的PCR引物再次擴(kuò)增上述患者FLT3基因,應(yīng)用illumina Miseq二代測(cè)序儀分析擴(kuò)增產(chǎn)物,并將測(cè)序結(jié)果與UCSC數(shù)據(jù)庫(kù)比對(duì)。結(jié)果:23例AML患者毛細(xì)管電泳檢測(cè)顯示,17例患者為1條ITD插入突變,3例患者為2條ITD插入突變,3例患者為3條ITD插入突變。高通量測(cè)序共檢測(cè)到的33條ITD中17條ITD插入序列為FLT3野生型完全重復(fù),16條ITD為野生型部分重復(fù)。1例伴多條ITD患者同一插入長(zhǎng)度存在兩種ITD序列,另外1例伴有2條ITD為同一ITD插入位點(diǎn)。ITD插入序列發(fā)生在p.Y572位至p.L602位之間,所有患者ITD均覆蓋p.V592-p.E598之間的一個(gè)或多個(gè)氨基酸。結(jié)論:Illumina Miseq二代測(cè)序儀可靈敏、準(zhǔn)確地分析FLT3-ITD序列,FLT3-ITD序列特征變化較大但突變熱點(diǎn)區(qū)域較為集中。
[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.

【作者單位】： 中日友好醫(yī)院檢驗(yàn)科;
【基金】：中日友好醫(yī)院青年課題(QN-2013-11)
【分類號(hào)】：R440;R733.71
【正文快照】： Fms樣酪氨酸激酶-3(fms-like tyrosine kinase-3,FLT3)是Ⅲ型受體酪氨酸激酶家族成員,在約20%-30%的急性髓系白血病(acute myeloid leuke-mia,AML)患者中FLT3基因中外顯子14和15存在內(nèi)部串聯(lián)重復(fù)(FLT3 internal tandem duplication, FLT3-ITD)[1]。FLT3-ITD對(duì)AML患者的完全緩，

本文編號(hào)：1388224