Establishment of a rapid and easy method for simultaneous detection of FLT3-ITD and NPM1 gene mutations in acute myeloid leukemia.
- Author:
Ying LU
1
;
Qiong WANG
;
Qi-tian MU
;
Meng-xia YU
;
Qin HUANG
;
Jie JIN
Author Information
- Publication Type:Journal Article
- MeSH: Base Sequence; Electrophoresis, Polyacrylamide Gel; methods; Female; Humans; Leukemia, Myeloid, Acute; genetics; Male; Molecular Sequence Data; Multiplex Polymerase Chain Reaction; methods; Mutation; Nuclear Proteins; genetics; fms-Like Tyrosine Kinase 3; genetics
- From: Chinese Journal of Medical Genetics 2012;29(2):163-166
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo establish a stable, rapid multiplex PCR assay combined with PAGE gel electrophoresis for simultaneously detecting FLT3-ITD and NPM1 mutations in acute myeloid leukemia (AML).
METHODSCapillary electrophoresis (CE) and PAGE gel electrophoresis were simultaneously used to analyze FLT3-ITD and NPM1 mutations in 117 de novo AML patients with normal cytogenetic findings.
RESULTSFor certain mutations, the length of mutated double-stranded DNA is longer than wild-type DNA. Since FLT3-mut (420 bp) is longer than FLT3-wt (327-332 bp), and NPM1-mut (172 bp) is longer than NPM1-wt (168 bp), heteroduplex will move more slowly during PAGE gel electrophoresis than homoduplex. Therefore the mutations may be detected. A total of 117 CN-AML patients were analyzed with CE and PAGE gel electrophoresis, and the results were identical, which included 18 (15.4%) patients with FLT3-ITD+/NPM1-, 19 (16.2%) patients with FLT3-ITD+/NPM1+, 25 (21.4%) patients with FLT3-ITD-/NPM1+, and 55 (47.0%) patients with FLT3-ITD-/NPM1-.
CONCLUSIONBoth types of electrophoresis assays may provide a rapid and handy assay for simultaneous detection of FLT3-ITD and NPM1 mutations. CE is relatively sensitive, stable; while PAGE electrophoresis is relatively simple, cheap, and reliable, which may be suitable for primary hospitals and preliminary screening.
