International Journal of Hematology

, Volume 107, Issue 2, pp 230–234 | Cite as

Quantification of Wilms’ tumor 1 mRNA by digital polymerase chain reaction

  • Yuki Koizumi
  • Daisuke Furuya
  • Teruo Endo
  • Kouichi Asanuma
  • Nozomi Yanagihara
  • Satoshi Takahashi
Original Article
  • 177 Downloads

Abstract

Wilms’ tumor 1 (WT1) is overexpressed in various hematopoietic tumors and widely used as a marker of minimal residual disease. WT1 mRNA has been analyzed using quantitative real-time polymerase chain reaction (real-time PCR). In the present study, we analyzed 40 peripheral blood and bone marrow samples obtained from cases of acute myeloid leukemia, acute lymphoblastic leukemia, and myelodysplastic syndrome at Sapporo Medical University Hospital from April 2012 to January 2015. We performed quantification of WT1 was performed using QuantStudio 3D Digital PCR System (Thermo Fisher Scientific‎) and compared the results between digital PCR and real-time PCR technology. The correlation between digital PCR and real-time PCR was very strong (R = 0.99), and the detection limits of the two methods were equivalent. Digital PCR was able to accurately detect lower WT levels compared with real-time PCR. Digital PCR technology can thus be utilized to predict WT1/ABL1 expression level accurately and should thus be useful for diagnosis or the evaluation of drug efficiency in patients with leukemia.

Keywords

WT1 ABL1 Minimal residual disease Digital PCR Real-time PCR 

Notes

Compliance with ethical standards

Conflict of interest

This research has no Grant numbers and sources of support. The authors declare no conflicts of interest.

References

  1. 1.
    Ogawa H, Tamaki H, Ikegame K, Soma T, Kawakami M, Tsuboi A, et al. The usefulness of monitoring WT1 gene transcripts for the prediction and management of relapse following allogeneic stem cell transplantation in acute type leukemia. Blood. 2003;101:1698–704.CrossRefPubMedGoogle Scholar
  2. 2.
    Cilloni D, Renneville A, Hermitte F, Hills RK, Daly S, Jovanovic JV, et al. Real-time quantitative polymerase chain reaction detection of minimal residual disease by standardized WT1 assay to enhance risk stratification in acute myeloid leukemia: a European LeukemiaNet study. J Clin Oncol. 2009;27:5195–201.CrossRefPubMedGoogle Scholar
  3. 3.
    Cheever MA, Allison JP, Ferris AS, Finn OJ, Hastings BM, Hecht TT, et al. The prioritization of cancer antigens: a national cancer institute pilot project for the acceleration of translational research. Clin Cancer Res. 2009;15:5323–37.CrossRefPubMedPubMedCentralGoogle Scholar
  4. 4.
    Hindson CM, Chevillet JR, Briggs HA, Gallichotte EN, Ruf IK, Hindson BJ, et al. Absolute quantification by droplet digital PCR versus analog real-time PCR. Nat Methods. 2013;10:1003–5.CrossRefPubMedPubMedCentralGoogle Scholar
  5. 5.
    Goh HG, Lin M, Fukushima T, Saglio G, Kim D, Choi SY, et al. Sensitive quantitation of minimal residual disease in chronic myeloid leukemia using nanofluidic digital polymerase chain reaction assay. Leuk Lymphoma. 2011;52:896–904.CrossRefPubMedGoogle Scholar
  6. 6.
    Jennings LJ, George D, Czech J, Yu M, Joseph L. Detection and quantification of BCR-ABL1 fusion transcripts by droplet digital PCR. J Mol Diagn. 2014;16:174–9.CrossRefPubMedGoogle Scholar
  7. 7.
    Iacobucci I, Lonetti A, Venturi C, Ferrari A, Papayannidis C, Ottaviani E, et al. Use of a high sensitive nanofluidic array for the detection of rare copies of BCR-ABL1 transcript in patients with Philadelphia-positive acute lymphoblastic leukemia in complete response. Leuk Res. 2014;38:581–5.CrossRefPubMedGoogle Scholar
  8. 8.
    Inoue K, Sugiyama H, Ogawa H, Nakagawa M, Yamagami T, Miwa H, et al. WT1 as a new prognostic factor and a new marker for the detection of minimal residual disease in acute leukemia. Blood. 1994;84:3071–9.PubMedGoogle Scholar
  9. 9.
    Van Dijk JP, Knops GH, Van De Locht LT, Menke AL, Jansen JH, Mensink EJ, et al. Abnormal WT1 expression in the CD34-negative compartment in myelodysplastic bone marrow. Br J Haematol. 2002;118:1027–33.CrossRefPubMedGoogle Scholar
  10. 10.
    Soverini S, Rosti G, Baccarani M, Martinelli G. Molecular monitoring. Curr Hematol Malig Rep. 2014;9:1–8.CrossRefPubMedGoogle Scholar
  11. 11.
    Nakamae H, Fujisawa S, Ogura M, Uchida T, Onishi Y, Taniwaki M, et al. Dasatinib versus imatinib in Japanese patients with newly diagnosed chronic phase chronic myeloid leukemia: a subanalysis of the DASISION 5-year final report. Int J Hematol. 2017;105:792–804.CrossRefPubMedGoogle Scholar
  12. 12.
    Kolostova K, Pinkas M, Jakabova A, Pospisilova E, Svobodova P, Spicka J, et al. Molecular characterization of circulating tumor cells in ovarian cancer. Am J Cancer Res. 2016;6:973–80.PubMedPubMedCentralGoogle Scholar

Copyright information

© The Japanese Society of Hematology 2017

Authors and Affiliations

  1. 1.Division of Laboratory MedicineSapporo Medical University HospitalSapporoJapan
  2. 2.Department of Infection Control and Laboratory MedicineSapporo Medical University School of MedicineSapporoJapan

Personalised recommendations