A comparison between cDNA and DNA samples for the detection of c-KIT exon 17 mutation in acute myeloid leukemia patients
10.3760/cma.j.issn.1009-8158.2019.06.011
- VernacularTitle:cDNA与DNA标本检测AML患者c-KIT基因外显子17突变的比较
- Author:
Lu YANG
1
;
Hong LIU
;
Wenmin CHEN
;
Lingdi LI
;
Lingyu LONG
;
Yanrong LIU
;
Yazhen QIN
Author Information
1. 北京大学人民医院北京大学血液病研究所
- Keywords:
Leukemia;
myeloid;
acute;
Proto-oncogene proteins c-kit;
Exons;
Mutation;
DNA;
Complementary;
DNA
- From:
Chinese Journal of Laboratory Medicine
2019;42(6):453-457
- CountryChina
- Language:Chinese
-
Abstract:
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=0.000073). (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=0.00003). (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.
