Detection of cytoplasmic nucleophosmin expression by imaging flow cytometry

Mutations within the nucleophosmin NPM1 gene occur in approximately one‐third of cases of acute myeloid leukemia (AML). These mutations result in cytoplasmic accumulation of the mutant NPM protein. NPM1 mutations are currently detected by molecular methods. Using samples from 37 AML patients, we inv...

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Veröffentlicht in:Cytometry. Part A 2012-10, Vol.81A (10), p.896-900
Hauptverfasser: Grimwade, Lizz, Gudgin, Emma, Bloxham, David, Bottley, Graham, Vassiliou, George, Huntly, Brian, Scott, Mike A., Erber, Wendy N.
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container_end_page 900
container_issue 10
container_start_page 896
container_title Cytometry. Part A
container_volume 81A
creator Grimwade, Lizz
Gudgin, Emma
Bloxham, David
Bottley, Graham
Vassiliou, George
Huntly, Brian
Scott, Mike A.
Erber, Wendy N.
description Mutations within the nucleophosmin NPM1 gene occur in approximately one‐third of cases of acute myeloid leukemia (AML). These mutations result in cytoplasmic accumulation of the mutant NPM protein. NPM1 mutations are currently detected by molecular methods. Using samples from 37 AML patients, we investigated whether imaging flow cytometry could be a viable alternative to this current technique. Bone marrow/peripheral blood cells were stained with anti‐NPM antibody and DRAQ5 nuclear stain, and data were acquired on an ImageStream imaging flow cytometer (Amnis Corp., Seattle, USA). Using the similarity feature for data analysis, we demonstrated that this technique could successfully identify cases of AML with a NPM1 mutation based on cytoplasmic NPM protein staining (at similarity threshold of 1.1 sensitivity 88% and specificity 90%). Combining data of mean fluorescence intensity and % dissimilar staining in a 0–2 scoring system further improved the sensitivity (100%). Imaging flow cytometry has the potential to be included as part of a standard flow cytometry antibody panel to identify potential NPM1 mutations as part of diagnosis and minimal residual disease monitoring. Imaging flow cytometry is an exciting technology that has many possible applications in the diagnosis of hematological malignancies, including the potential to integrate modalities. © 2012 International Society for Advancement of Cytometry.
doi_str_mv 10.1002/cyto.a.22116
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These mutations result in cytoplasmic accumulation of the mutant NPM protein. NPM1 mutations are currently detected by molecular methods. Using samples from 37 AML patients, we investigated whether imaging flow cytometry could be a viable alternative to this current technique. Bone marrow/peripheral blood cells were stained with anti‐NPM antibody and DRAQ5 nuclear stain, and data were acquired on an ImageStream imaging flow cytometer (Amnis Corp., Seattle, USA). Using the similarity feature for data analysis, we demonstrated that this technique could successfully identify cases of AML with a NPM1 mutation based on cytoplasmic NPM protein staining (at similarity threshold of 1.1 sensitivity 88% and specificity 90%). Combining data of mean fluorescence intensity and % dissimilar staining in a 0–2 scoring system further improved the sensitivity (100%). 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Combining data of mean fluorescence intensity and % dissimilar staining in a 0–2 scoring system further improved the sensitivity (100%). Imaging flow cytometry has the potential to be included as part of a standard flow cytometry antibody panel to identify potential NPM1 mutations as part of diagnosis and minimal residual disease monitoring. 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source MEDLINE; Wiley Online Library Journals Frontfile Complete; Elektronische Zeitschriftenbibliothek - Frei zugängliche E-Journals; Wiley Free Content; Alma/SFX Local Collection
subjects acute leukemia
Acute myeloid leukemia
Adult
Aged
Aged, 80 and over
Anthraquinones
Antibodies
Bone Marrow - metabolism
Bone Marrow - pathology
Cell Nucleus - genetics
Cell Nucleus - metabolism
Cytoplasm - genetics
Cytoplasm - metabolism
Female
Flow Cytometry
Gene Expression
hematology
Humans
imaging flow cytometry
Leukemia, Myeloid, Acute - diagnosis
Leukemia, Myeloid, Acute - genetics
Leukemia, Myeloid, Acute - metabolism
Leukocytes, Mononuclear - metabolism
Leukocytes, Mononuclear - pathology
Male
Middle Aged
Mutation
Neoplasm, Residual - diagnosis
Neoplasm, Residual - genetics
Neoplasm, Residual - metabolism
Nuclear Proteins - genetics
Nuclear Proteins - metabolism
nucleophosmin
Research Design
Sensitivity and Specificity
Staining and Labeling
title Detection of cytoplasmic nucleophosmin expression by imaging flow cytometry
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