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DHPLC技术在白血病基因检测应用专题
A denaturing high-performance
liquid chromatography (DHPLC) assay could be used to identify NPM1
mutations in leukemia
11-14-2006
According to recent research from Italy, "NPM1 gene
mutations are the most frequent genetic lesion in the 60% of adult
acute myeloid leukemias (AMLs) with normal karyotype and no evidence of typical fusion genes
(BCR/ABL1, PML/RARA, AML1/ETO, CBFB/MYH11, DEK/CAN). Using direct
sequencing we previously identified six different heterozygous mutants
within exon 12 encoding the nucleophosmin C-terminus."
"Because of these mutations the shuttling protein nucleophosmin is aberrantly delocalized in the
cytoplasm of leukemic cells (NPMc+),"
explained G. Roti and colleagues, University
of Perugia. "Here, we designed and tested a DHPLC assay to detect
NPM1 mutated variants. To assess specificity, sensitivity, reliability,
and reproducibility, we analyzed DNA from 120 primary adult AMLs and
compared DHPLC results with immunohistochemistry
and sequencing.
"All electropherogram profiles in the 26
NPMc+ leukemias
were different from the wild type, indicating 100% sensitivity.
Sequencing categorized mutations A, B, and D, and all mutation A cases gave identical elution profiles. The other
mutations showed typical chromatograms, with mutations B and D differing
for one nucleotide. Elution profiles and sequencing also identified
four new variants."
The researchers concluded, "Our results suggest that DHPLC detects
NPM1 mutations as well as direct sequencing and immunohistochemistry,
providing a helpful approach in the diagnosis of NPMc+
AML."
Roti and colleagues published their study in
the Journal of Molecular Diagnostics (Denaturing high-performance
liquid chromatography - A valid approach for identifying NPM1 mutations
in acute myeloid leukemia. J Mol Diagn, 2006;8(2):254-259).
For additional information, contact C. Mecucci,
University of Perugia, Laboratory of Cytogenetics
and Molecular Genetics, I-06123 Perugia, Italy.
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