Detection of NPM1 Mutation in Acute Myeloid Leukemia by Droplet Digital PCR and Its Clinical Application Value.
10.19746/j.cnki.issn.1009-2137.2022.03.001
- Author:
Ye JIN
1
;
Shi Sen WANG
1
;
Pei Hui XIA
2
;
Qian YUAN
1
;
Gao Fei XIAO
2
;
Jiang LIN
2
;
Jia Yan LENG
1
;
Yu Juan MA
1
;
Jun QIAN
3
Author Information
1. Department of Hematology, The Affiliated People's Hospital of Jiangsu University,Zhenjiang 212002, Jiangsu Province, China；Zhenjiang Clinical Research Center of Hematology,Zhenjiang 212002, Jiangsu Province, China.
2. Zhenjiang Clinical Research Center of Hematology,Zhenjiang 212002, Jiangsu Province, China；Laboratory Center, The Affiliated People's Hospital of Jiangsu University, Zhenjiang 212002, Jiangsu Province, China.
3. Department of Hematology, The Affiliated People's Hospital of Jiangsu University,Zhenjiang 212002, Jiangsu Province, China；Zhenjiang Clinical Research Center of Hematology,Zhenjiang 212002, Jiangsu Province, China,Email: qianjun0007@hotmail.com.
- Publication Type:Journal Article
- Keywords:
Droplet digital PCR;
NPM1;
acute myeloid leukemia;
minimal residual disease
- MeSH:
Humans;
Leukemia, Myeloid, Acute/therapy*;
Mutation;
Nuclear Proteins/genetics*;
Nucleophosmin;
Polymerase Chain Reaction;
Prognosis
- From:
Journal of Experimental Hematology
2022;30(3):653-658
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVE:To establish the droplet digital PCR (ddPCR) assay for the detection of NPM1 type A mutation in patients with acute myeloid leukemia (AML), and to evaluate its specificity, sensitivity and its value in clinical application.
METHODS:NPM1 mutant and wildtype plasmids were used to verify the performance of ddPCR. Both ddPCR and Sanger sequencing were used to detect the bone marrow samples of 87 AML patients, which were confirmed by next generation sequencing (NGS). Moreover, NPM1 mutation burden was dynamically monitored in five patients by ddPCR.
RESULTS:The limit of blank (LOB) of ddPCR established for NPM1 mutation detection was 1.1 copies/μl, and the limit of detection (LOD) was 2.43 copies/μl, which had good linearity. Among the 87 newly diagnosed AML patients, ddPCR identified seventeen cases positive for NPM1 mutation (19.5%)， which was consistent with Sanger sequencing. NGS confirmed 12 positive cases, including 8 of type A mutations, 2 of type D mutations, and 2 of rare type mutations. The results of dynamic monitoring of NPM1 mutation burden in 5 patients showed that the NPM1 mutation burden decreased obviously even close to 0, when patients achieve complete remission after chemotherapy. However, the mutation burden was increased again at the time of relapse.
CONCLUSION:In this study, we established a ddPCR method for detection of NPM1 mutation with good sensitivity and repeatability, which can be used for screening NPM1 mutation in newly diagnosed AML patients and for minimal residual disease monitoring after remission in positive AML patients to guide treatment.
