Performance and Clinical Evaluation of a Sensitive Multiplex Assay for the Rapid Detection of Common NPM1 Mutations Michael Hafez, Fei Ye, Keith Jackson,

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Presentation transcript:

Performance and Clinical Evaluation of a Sensitive Multiplex Assay for the Rapid Detection of Common NPM1 Mutations Michael Hafez, Fei Ye, Keith Jackson, Zhe Yang, Judith E. Karp, Emmanuel Labourier, Christopher D. Gocke The Journal of Molecular Diagnostics Volume 12, Issue 5, Pages 629-635 (September 2010) DOI: 10.2353/jmoldx.2010.090219 Copyright © 2010 American Society for Investigative Pathology and Association for Molecular Pathology Terms and Conditions

Figure 1 Analytical performance evaluation. A: Wild-type and mutant sequences detected by the assay. Bold indicates insertion mutations. The relative representation of each mutation in the total mutation-positive population is indicated.1617 B: Representative examples of signal (MFI) obtained with a serial dilution of a mutant A-positive cell line RNA (OCI-AML3) in wild-type cell line RNA (HL-60). C: Representative examples of signal (MFI) obtained with a serial dilution of mutant A in vitro transcript (mut-A) in wild-type cell line RNA (HL-60). D: Representative examples of data obtained with a serial dilution of four different in vitro transcripts (mut-A, mut-B, mut-D, or mut-J) in wild-type cell line RNA (HL-60). The graph shows the ratio of mutant to wild-type signal according to the initial copy number input of each in vitro transcript. WT, wild-type. The Journal of Molecular Diagnostics 2010 12, 629-635DOI: (10.2353/jmoldx.2010.090219) Copyright © 2010 American Society for Investigative Pathology and Association for Molecular Pathology Terms and Conditions

Figure 2 Comparison with a size-based assay. A: Summary of the positive (Pos) and negative (Neg) calls obtained with serial dilutions of a mutant A-positive cell line RNA (OCI-AML3) or in vitro transcript (mut-A) in wild-type cell line RNA (HL-60). A Pos/Neg call indicates that the results were not reproducible (below or at the limit of detection of the method). B: Representative examples of capillary electrophoresis traces. The wild-type (WT)- and mutant (M)-specific peaks are indicated. The small peaks at the beginning and end of each trace correspond to molecular weight markers. The Journal of Molecular Diagnostics 2010 12, 629-635DOI: (10.2353/jmoldx.2010.090219) Copyright © 2010 American Society for Investigative Pathology and Association for Molecular Pathology Terms and Conditions