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Rapid and sensitive typing of NPM1 mutations using LNA-mediated PCR clamping

Leukemia volume 20, pages 1897–1899 (2006)Cite this article

The actual detection level is clearly lower than achievable with other procedures,6 and thus might be insufficient to perform MRD analysis with high sensitivity, detecting cell populations far below 1/1000 cells. However, the flexibility of the assay makes it especially suitable for rapid assessment of the presence of mutations in follow-up samples. This is of major importance in those cases in which only material after start of treatment is available, or when the precise type of mutation is unknown, because initial diagnosis and MRD-assessment were performed in different laboratories. In addition, this assay might help to identify the precise mutation type present in those 20% not having mutation Type A, especially for the design of real-time PCR primers and probe sets for MRD quantification.6

Taken together, the simple procedure described here enables the rapid assessment of NPM1 mutations with a sensitivity of 1/100 to 1/1000 and avoids the necessity of cloning or other steps to unambiguously identify NPM1 alterations. The assay was able to detect more than 30 different NPM1 mutations, thus seems to work with the vast majority of mutations reported so far. It appears to represent a versatile tool for NPM1 mutation detection and might be combined with other assays like fragment analysis or real-time PCR to improve MRD detection. This is especially relevant in those cases having not one of the typical mutations (Type A, B, D), where establishing an individual real-time PCR assay might be too time consuming in the clinical situation.

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Authors and Affiliations

  1. Medizinische Klinik und Poliklinik I, Universitätsklinikum Carl Gustav Carus der Technischen Universität, Dresden, Germany

    C Thiede, E Creutzig, T Illmer, M Schaich & G Ehninger

  2. CHIPRON GmbH, Berlin, Germany

    V Heise

  3. TIB MOLBIOL GmbH, Berlin, Germany

    O Landt

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  1. C Thiede
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  2. E Creutzig
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  3. T Illmer
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Correspondence to C Thiede.

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Thiede, C., Creutzig, E., Illmer, T. et al. Rapid and sensitive typing of NPM1 mutations using LNA-mediated PCR clamping. Leukemia 20, 1897–1899 (2006). https://doi.org/10.1038/sj.leu.2404367

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