2008
DOI: 10.2353/jmoldx.2008.070166
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An Allele-Specific RT-PCR Assay to Detect Type A Mutation of the Nucleophosmin-1 Gene in Acute Myeloid Leukemia

Abstract: Nucleophosmin-1 (NPM1) mutations represent the most frequent gene alteration in acute myeloid leukemia (AML). The most common NPM1 mutation type, accounting for 75 to 80% of cases , is referred to as mutation A (NPM1-mutA). These NPM1 alterations have been shown to possess prognostic significance because they appear to identify patients who will benefit from chemotherapy. Given the high prevalence and stability of these mutations over the course of disease , NPM1 mutations may serve as ideal targets for minima… Show more

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Cited by 36 publications
(25 citation statements)
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“…Correspondence: Pier Giuseppe Pelicci, European Institute of Oncology at the IFOM-IEO Campus, Via Adamello, 16, 20139 Milan, Italy; e-mail: piergiuseppe.pelicci@ifom-ieo-campus.it.…”
Section: Acknowledgmentsmentioning
confidence: 99%
See 1 more Smart Citation
“…Correspondence: Pier Giuseppe Pelicci, European Institute of Oncology at the IFOM-IEO Campus, Via Adamello, 16, 20139 Milan, Italy; e-mail: piergiuseppe.pelicci@ifom-ieo-campus.it.…”
Section: Acknowledgmentsmentioning
confidence: 99%
“…7 Currently available methods to detect NPM1 mutations include DNA sequencing, real-time quantitative polymerase chain reaction (PCR), denaturing high-performance liquid chromatography, capillary electrophoresis, locked nucleic acid-mediated PCR clamping, and allele-specific reverse-transcribed PCR assays. [10][11][12][13][14][15][16][17][18] Although highly specific, these methods are expensive and laborious. Alternatively, NPM1 mutations can be identified indirectly, using antibodies that recognize epitopes that are common for both the wild-type (wt) and mutant NPM1 proteins and allow recognition of the cytoplasmic dislocation of NPM1.…”
Section: Introductionmentioning
confidence: 99%
“…23 A number of methods have been developed to rapidly detect NPM1 mutations in genomic DNA and/or mRNA specimens. 7,17,[24][25][26][27] The most widely adopted screening method, routinely used in our laboratory, is polymerase chain reaction (PCR) amplification of the mutational hotspot region followed by capillary electrophoresis analysis. This approach yields semiquantitative results.…”
mentioning
confidence: 99%
“…Recently, several groups have designed quantitative real-time polymerase chain reaction (qPCR)-based assays for detecting single or multiple common NPM1 mutations in AML. [25][26][27] These methods offer the advantage of reliable quantification of mutated NPM1 and, therefore, would be useful for providing a quantitative measurement of disease burden and for assessing MRD after therapy. [25][26][27] The clinical applications of these assays, however, and whether they are ready to replace conventional capillary electrophoresis assays are still being debated.…”
mentioning
confidence: 99%
“…In addition, the sensitivity of this method is low, with the lower limit of detection beginning at ~20% (10). Therefore, polymerase chain reaction (PCR) methods such as electrophoresis (11), melting curve analysis (12), high-resolution melting (HRM) analysis which detects the different melting temperatures of the PCR products (13), locked nucleic acid clamp-mediated PCR (10,14), and capillary electrophoresis (15,16) have been investigated as potentially more sensitive methods for detection of this 4 bp mutation.…”
Section: Introductionmentioning
confidence: 99%