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Article

Establishment of a quenching probe method for detection of NPM1 mutations in acute myeloid leukemia cells

  • Authors:
    • Noriko Kawaguchi‑Ihara
    • Mai Itoh
    • Ikuo Murohashi
    • Shuji Tohda
  • View Affiliations / Copyright

    Affiliations: Department of Laboratory Medicine, Tokyo Medical and Dental University, Bunkyo‑ku, Tokyo 113‑8519, Japan, Department of Health Sciences, Saitama Prefectural University, Koshigaya, Saitama 343‑8540, Japan
  • Pages: 2429-2432
    |
    Published online on: February 11, 2016
       https://doi.org/10.3892/ol.2016.4225
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Abstract

Nucleophosmin (NPM1) mutations, generally consisting of a four base-pair insertion, are present in ~60% of all cytogenetically normal acute myeloid leukemia (AML) cases. The mutation is clinically significant as an important prognostic factor. Direct sequencing is the current standard method of mutation detection, however, it is quite costly and time consuming. The present study aimed to establish a highly sensitive quenching probe (QP) method to detect NPM1 mutations efficiently. Melting curve analysis was performed using a QP, following polymerase chain reaction for amplification of the involved region of the gene. The curve derived from the fluorescent intensity with respect to the temperature of OCI/AML3, a heterozygous NPM1 mutant AML cell line, was W‑shaped with melting peaks at 61˚C and 68˚C. That of M‑07e, the homozygous wild type cell line, was V‑shaped with a melting peak at 68˚C. Thus, the curve derived from the mutant allele was easily discriminated from that of the wild‑type allele. The mutant allele was detected in concentrations as low as 3% as determined by a subsequent sensitivity study. With a short testing time and a high sensitivity, this assay was applicable for NPM1‑mutated AML patient samples and is appropriate for screening NPM1 mutations. It does require further examination as to whether it would be useful as a detection method for other mutant alleles since NPM1 mutations may consist of 61 known types of mutant sequences. To the best of our knowledge, this is the first report describing the QP method for the detection of NPM1 mutations.
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Copy and paste a formatted citation
Spandidos Publications style
Kawaguchi‑Ihara N, Itoh M, Murohashi I and Tohda S: Establishment of a quenching probe method for detection of NPM1 mutations in acute myeloid leukemia cells. Oncol Lett 11: 2429-2432, 2016.
APA
Kawaguchi‑Ihara, N., Itoh, M., Murohashi, I., & Tohda, S. (2016). Establishment of a quenching probe method for detection of NPM1 mutations in acute myeloid leukemia cells. Oncology Letters, 11, 2429-2432. https://doi.org/10.3892/ol.2016.4225
MLA
Kawaguchi‑Ihara, N., Itoh, M., Murohashi, I., Tohda, S."Establishment of a quenching probe method for detection of NPM1 mutations in acute myeloid leukemia cells". Oncology Letters 11.4 (2016): 2429-2432.
Chicago
Kawaguchi‑Ihara, N., Itoh, M., Murohashi, I., Tohda, S."Establishment of a quenching probe method for detection of NPM1 mutations in acute myeloid leukemia cells". Oncology Letters 11, no. 4 (2016): 2429-2432. https://doi.org/10.3892/ol.2016.4225
Copy and paste a formatted citation
x
Spandidos Publications style
Kawaguchi‑Ihara N, Itoh M, Murohashi I and Tohda S: Establishment of a quenching probe method for detection of NPM1 mutations in acute myeloid leukemia cells. Oncol Lett 11: 2429-2432, 2016.
APA
Kawaguchi‑Ihara, N., Itoh, M., Murohashi, I., & Tohda, S. (2016). Establishment of a quenching probe method for detection of NPM1 mutations in acute myeloid leukemia cells. Oncology Letters, 11, 2429-2432. https://doi.org/10.3892/ol.2016.4225
MLA
Kawaguchi‑Ihara, N., Itoh, M., Murohashi, I., Tohda, S."Establishment of a quenching probe method for detection of NPM1 mutations in acute myeloid leukemia cells". Oncology Letters 11.4 (2016): 2429-2432.
Chicago
Kawaguchi‑Ihara, N., Itoh, M., Murohashi, I., Tohda, S."Establishment of a quenching probe method for detection of NPM1 mutations in acute myeloid leukemia cells". Oncology Letters 11, no. 4 (2016): 2429-2432. https://doi.org/10.3892/ol.2016.4225
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