cDNA与DNA标本检测AML患者c-KIT基因外显子17突变的比较  

A comparison between cDNA and DNA samples for the detection of c-KIT exon 17 mutation in acute myeloid leukemia patients

作　　者：杨璐[1] 刘虹[2] 陈文敏 李玲娣[1] 龙玲玉 刘艳荣[1] 秦亚溱[1] Yang Lu;Liu Hong;Chen Wenmin;Li Lingdi;Long Lingyu;Liu Yanrong;Qin Yazhen(Institute of Hematology, Peking University Peoples′ Hospital, Beijing 100044, China;Xinjiang Uygur Autonomous Region People′s Hospital, Uygur 830001, China)

机构地区：[1]北京大学人民医院北京大学血液病研究所,北京100044 [2]新疆维吾尔自治区人民医院血液科,乌鲁木齐830001

出　　处：《中华检验医学杂志》2019年第6期453-457,共5页Chinese Journal of Laboratory Medicine

基　　金：国家自然科学基金(81570130、81870125).

摘　　要：目的研究标本类型对急性髓系白血病(AML)患者c-KIT基因外显子17突变检测结果的影响。方法回顾性研究。收集2016年6月至2018年8月送检北京大学人民医院的骨髓标本共51份,采集自37例AML患者初诊或治疗后,其中男17例,女20例,送检时中位年龄33岁(1~82岁)。t(8;21)AML24例,inv(16)/t(16;16)AML11例,非CBF-AML2例。标本同时提取RNA和DNA,采用Sanger测序法分别检测cDNA及DNA标本的c-KIT外显子17突变,比较配对标本的检测结果。结果(1)51对标本中14对均检出c-KIT突变,17对均未检出突变,其余20对均为cDNA检出而DNA未检出突变,不一致率为39.2%,cDNA突变检出率明显高于DNA标本(66.7%vs27.5%,χ2=15.74,P=0.000073)。(2)t(8;21)AML、inv(16)AML及非CBF-AML患者的配对标本均存在突变结果不一致现象,不一致率分别为36.4%(12/33)、27.2%(3/11)和71.4%(5/7)。(3)治疗后标本的不一致率明显高于初诊标本(72.7%vs13.8%,χ2=18.22,P=0.00003)。(4)5例具有c-KIT突变患者治疗随访中均可见cDNA与DNA标本结果不一致现象。结论对于Sanger测序法检测AML患者c-KIT外显子17突变,cDNA比DNA标本具有更高的检出率,在治疗后尤其普遍。Objective To investigate the impact of sample typeon the detection of c-KIT exon 17 mutation in acute myeloid leukemia (AML) patients. Methods A retrospective study was conducted on 51 bone marrow samples collected from 37 AML patients [17 maleand 20 female, with a median age of 33 (range from 1 to 82)] at diagnosis or after treatment from June 2016 to August 2018. Of the 37 cases of AML, 24 were t(8;21) AML, 11 were inv(16)/t(16;16) AML and 2 were non-CBF-AML. RNA and DNA were simultaneously extracted from every sample. PCR followed by Sanger sequencing were used to screen c-KIT exon 17 mutation, and the comparisons were made between paired cDNA and DNAsamples. Results (1) Of the 51 paired samples, 14 pairs were simultaneously detected positive for c-KITmutation in both of cDNA and DNA samples, but 17 pairs were detected negative in both, and the remaining 20 pairswere only detected positive for the mutation in cDNA but not in DNA, with an inconsistency rate of 39.2%. The positive rate of detecting c-KITmutation was significantly higher in cDNA than in DNA samples (66.7% vs 27.5%,P=0.000 073).(2)Inconsistent mutation results between paired cDNA and DNA samples occurred in t(8;21)AML, inv(16)AML and non-CBF-AML patients with the inconsistency rate of 36.4%(12/33), 27.2%(3/11) and 71.4%(5/7), respectively.(3)The inconsistency rate was significantly higher in samples collected after treatment compared with those collected at diagnosis (72.7% vs 13.8%, P=0.000 03).(4) All 5 serially monitored patients with c-KITmutation had inconsistency in mutation detection between cDNA and DNA samples during follow up. Conclusion cDNA improves the detection of c-KIT exon 17 mutation in AML patients compared with DNA, which is especially common after treatment.

关 键 词：白血病 髓样 急性 原癌基因蛋白质类c-kit 外显子 突变 DNA 互补 DNA 

分 类 号：R733.71[医药卫生—肿瘤] R730.43[医药卫生—临床医学]