Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Letter to the Editor
  • Published:

MassARRAY assay: a more accurate method for JAK2V617F mutation detection in Chinese patients with myeloproliferative disorders

Leukemia volume 22, pages 660–663 (2008)Cite this article

The JAK2V617F mutation has recently been described as an essential oncogenetic event associated with the myeloproliferative disorders (MPD).1 Several techniques have emerged to detect this kind of mutation. As a newly developed one, MassARRAY assay combines competitive PCR with MALDI-TOF (matrix-assisted laser desorption/ionization time-of-flight) mass spectrometry and is an advanced system for automated high-throughput analysis of mutations or single nucleotide polymorphisms (SNPs).2 Other methods of SNPs detection, which previously reported suitable for JAK2V617F mutation, included restriction fragment length polymorphism (RFLP),3 direct sequencing analysis,4 pyrosequencing,5 real-time AS-PCR6 and a denaturing high-performance liquid chromatography (DHPLC) assay.7 RFLP analysis is the simplest and cheapest method, not requiring specific instruments. Although it can be used as the first screening method for JAK2V617F mutation, there may be false positives because of incomplete digestion,3 and positive results need to be confirmed by another detection system. Direct sequencing analysis is a golden rule of SNP detection with high specificity and can identify any unknown SNP, providing detailed information and ‘direct visualization’ of sequence information. However, the sensitivity of sequencing is rather low (>5%), its laborious, time-consuming and expensive procedure needs no mention.4 Pyrosequencing is a quantitative and rapid method, presenting sequence data ‘in real time’ without post-PCR procession. But it is also not sensitive enough (5–10%).5 Real-time AS-PCR is a more sensitive method (0.1%) for allele-specific real-time quantification, while the design of the primer and probe is complex and each lab needs an individual cutoff point for analyzing any SNPs, otherwise the false positives might occur.6 The DHPLC assay also needs post-PCR procession, thus elaborating and technically challenging.7

Among the clinical features, no differences of sex and age were found between MPD patients positive and negative for JAK2V617F mutation. Interestingly, comparing with wild-type cases, PV patients with homozygous mutation presented a significant elevation of white blood cell counts (30.8±8.0 × 109 l−1 vs 9.0±1.4 × 109 l−1, P=0.005). Moreover, higher hemoglobin concentration was found in the cases with heterozygous mutation (183.1±5.3 g l−1) than those with wild-type (150.2±15.7 g l−1, P=0.014). In vitro, erythropoietin hypersensitivity was induced in murine hematopoietic cells transduced with JAK2V617F.8 In vivo, expression of JAK2V617F in a murine bone marrow transplant model developed features of PV.8 Therefore, this mutation could cause abnormal proliferation of hematopoietic progenitors and related to PV pathogenesis.

This is a preview of subscription content, access via your institution

Relevant articles

Open Access articles citing this article.

Access options

Buy this article

  • Purchase on Springer Link
  • Instant access to full article PDF
Buy now

Prices may be subject to local taxes which are calculated during checkout

Figure 1
Figure 2

References

  1. Delaval B, Lelièvre H, Birnbaum D . Myeloproliferative disorders: the centrosome connection. Leukemia 2005; 19: 1739–1744.

    Article  CAS  Google Scholar 

  2. Shi JY, Shi ZZ, Zhang ZJ, Zhu YM, Gu BW, Li G et al. Association between single nucleotide polymorphisms in deoxycytidine kinase and treatment response among acute myeloid leukaemia patients. Pharmacogenetics 2004; 14: 759–768.

    Article  CAS  Google Scholar 

  3. Cantor CR, Campbell PJ, Scott LM, Buck G, Wheatley K, East CL et al. Definition of subtypes of essential thrombocythaemia and relation to polycythaemia vera based on JAK2 V617F mutation status: a prospective study. Lancet 2005; 366: 1945–1953.

    Article  Google Scholar 

  4. James C, Delhommeau F, Marzac C, Teyssandier I, Couedic JP, Giraudier S et al. Detection of JAK2 V617F as a first intention diagnostic test for erythrocytosis. Leukemia 2006; 20: 350–353.

    Article  CAS  Google Scholar 

  5. Jones AV, Kreil S, Zoi K, Waghorn K, Curtis C, Zhang L et al. Widespread occurrence of the JAK2 V617F mutation in chronic myeloproliferative disorders. Blood 2005; 106: 2162–2168.

    Article  CAS  Google Scholar 

  6. Nussenzveig RH, Swierczek SI, Jelinek J, Gaikwad A, Enli L, Verstovsek S et al. Polycythemia vera is not initiated by JAK2V617F mutation. Exp Hematol 2007; 35: 32–38.

    Article  CAS  Google Scholar 

  7. Sattler M, Walz C, Crowley BJ, Lengfelder E, Janne PA, Rogers AM et al. A sensitive high-throughput method to detect activating mutations of Jak2 in peripheral-blood samples. Blood 2006; 107: 1237–1238.

    Article  CAS  Google Scholar 

  8. Lacout C, Pisani DF, Tulliez M, Gachelin FM, Vainchenker W, Villeval JL . JAK2V617F expression in murine hematopoietic cells leads to MPD mimicking human PV with secondary myelofibrosis. Blood 2006; 108: 1652–1660.

    Article  CAS  Google Scholar 

Download references

Acknowledgements

This work was supported, in part, by the Chinese National Key Program for Basic Research (973:2004CB518600), the Chinese National High Tech Program (863:2006AA02A405 and 863:2006AA02A301), the National Natural Science Foundation of China (30521003), the Key Discipline Program of Shanghai Municipal Education Commission (Y0201), the Shanghai Commission of Science and Technology (06DZ22021), the Shanghai Rising Star Program (05QMX1429), the Scientific Research Foundation for the Returned Overseas Chinese Scholars and by the Samuel Waxman Cancer Research Foundation Laboratory, the Co-PI Program of Shanghai Rui Jin Hospital/Shanghai Jiao Tong University School of Medicine.

Author information

Author notes

  1. J-F Fu, J-Y Shi, W-L Zhao and G Li: These authors contributed equally to this work

Authors and Affiliations

  1. State Key Laboratory of Medical Genomics, Shanghai Institute of Hematology, Shanghai Rui Jin Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, China

    J-F Fu, J-Y Shi, W-L Zhao, G Li, Q Pan, J-M Li, J Hu, Z-X Shen & S-J Chen

  2. Zhejiang Institute of Hematology, The First Affiliated Hospital, Medical School of Zhejiang University, Hangzhou, China

    J Jin

  3. Department of Hematology, Shanghai Ren Ji Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, China

    F-Y Chen

Authors
  1. J-F Fu
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. J-Y Shi
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. W-L Zhao
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. G Li
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Q Pan
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. J-M Li
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. J Hu
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Z-X Shen
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. J Jin
    View author publications

    You can also search for this author in PubMed Google Scholar

  10. F-Y Chen
    View author publications

    You can also search for this author in PubMed Google Scholar

  11. S-J Chen
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Fu, JF., Shi, JY., Zhao, WL. et al. MassARRAY assay: a more accurate method for JAK2V617F mutation detection in Chinese patients with myeloproliferative disorders. Leukemia 22, 660–663 (2008). https://doi.org/10.1038/sj.leu.2404931

Download citation

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/sj.leu.2404931

This article is cited by

Search

Advanced search

Quick links