title: Detection of mutant NPM1 mRNA in acute myeloid leukemia using custom gene expression arrays.
abstract: Mutations in the gene encoding nucleophosmin (NPM1) carry a prognostic value for patients with acute myeloid leukemia (AML). Various techniques are currently being used to detect these mutations in routine molecular diagnostics. Incorporation of accurate NPM1 mutation detection on a gene expression platform would enable simultaneous detection with various other expression biomarkers. Here we present an array-based mutation detection using custom probes for NPM1 WT mRNA and NPM1 type A, B, and D mutant mRNA. This method was 100% accurate on a training cohort of 505 newly diagnosed unselected AML cases. Validation on an independent cohort of 143 normal-karyotype AML cases revealed no false-negative results, and one false positive (sensitivity 100.0% and specificity 98.7%). Based on this, we conclude that this method provides a reliable method for NPM1 mutation detection. The method can be applied to other genes/mutations as long as the mutant alleles are sufficiently highly expressed.
authors.affiliation: Skyline Diagnostics BV, Rotterdam, The Netherlands.
authors.forename: Martin H
authors.initials: MH
authors.lastname: van Vliet
authors.forename: Belinda
authors.initials: B
authors.lastname: Dumee
authors.forename: Erik
authors.initials: E
authors.lastname: Simons
authors.forename: Lars
authors.initials: L
authors.lastname: Bullinger
authors.forename: Konstanze
authors.initials: K
authors.lastname: Döhner
authors.forename: Hartmut
authors.initials: H
authors.lastname: Döhner
authors.forename: Henk
authors.initials: H
authors.lastname: Viëtor
authors.forename: Bob
authors.initials: B
authors.lastname: Löwenberg
authors.forename: Peter J M
authors.initials: PJ
authors.lastname: Valk
authors.forename: Erik H
authors.initials: EH
authors.lastname: van Beers
mesh_terms: D000465:Algorithms; D000483:Alleles; D015342:DNA Probes; D005260:Female; D015870:Gene Expression; D006801:Humans; D015470:Leukemia, Myeloid, Acute / Q000235:genetics* / Q000378:metabolism; D008297:Male; D009154:Mutation; D009687:Nuclear Proteins / Q000235:genetics* / Q000378:metabolism*; D000090243:Nucleophosmin; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods*; D011379:Prognosis; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D012680:Sensitivity and Specificity
chemical_list: D015342:DNA Probes; C000716967:NPM1 protein, human; D009687:Nuclear Proteins; D012333:RNA, Messenger; D000090243:Nucleophosmin