Evaluation of methods for detection of NPM1 gene mutations in acute myeloid leukemia.
- Author:
Zhi-Peng LI
1
;
Xuan ZHANG
;
Xiao-Ming ZHAO
;
Qing-Ge LI
;
Ya-Mei CHEN
;
Quan-Yi LU
Author Information
1. School of Life Science, Xiamen University, Xiamen 361005, Fujian Province, China.
- Publication Type:Journal Article
- MeSH:
Alleles;
DNA Mutational Analysis;
Electrophoresis, Capillary;
methods;
Genome;
Humans;
Leukemia, Myeloid, Acute;
diagnosis;
genetics;
Mutation;
Neoplasm, Residual;
diagnosis;
Nuclear Proteins;
genetics;
Plasmids;
Polymerase Chain Reaction;
methods;
Sensitivity and Specificity
- From:
Journal of Experimental Hematology
2011;19(4):999-1004
- CountryChina
- Language:Chinese
-
Abstract:
The purpose of this study was to establish real-time based methods for detection of NPM1 gene mutation in acute myeloid leukemia (AML). Primers/probes were designed according to the clustered region of NPM1 mutations on exon 12. Two real-time PCR assays, including high resolution melting curve (HRM) and allele-specific PCR (AS-PCR), were developed and clinically evaluated with 89 AML samples, which were parallelly detected by capillary electrophoresis (CE) and sequencing. The results showed that a total of 17 mutation-positive samples were detected, including type A (15 cases), type B (1 case) and type Nm (1 case). HRM assay could detect all mutant types, and the analytical sensitivity was around 5%. In contrast, AS-PCR assay detected only 95% mutant types, but its sensitivity was as high as 0.01%. It is concluded that considering the characteristics of each method as well as the clinical evaluation results, HRM may be used for screening of NPM1 mutations at diagnosis, while the AS-PCR can be used for the MRD quantification during follow-up.
