Clonality in myeloproliferative disorders: analysis by means of the polymerase chain reaction.

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  • D G Gilliland
    Department of Medicine, Brigham and Women's Hospital, Harvard Medical School, Boston, MA 02115.
  • K L Blanchard
    Department of Medicine, Brigham and Women's Hospital, Harvard Medical School, Boston, MA 02115.
  • J Levy
    Department of Medicine, Brigham and Women's Hospital, Harvard Medical School, Boston, MA 02115.
  • S Perrin
    Department of Medicine, Brigham and Women's Hospital, Harvard Medical School, Boston, MA 02115.
  • H F Bunn
    Department of Medicine, Brigham and Women's Hospital, Harvard Medical School, Boston, MA 02115.

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Abstract

<jats:p>The myeloproliferative syndromes are acquired disorders of hematopoiesis that provide insights into the transition from somatic cell mutation to neoplasia. The clonal origin of specific blood cells can be assessed in patients with X chromosome-linked polymorphisms, taking advantage of random inactivation of the X chromosome. We have adapted the PCR for determination of clonality on as few as 100 cells, including individual colonies grown in culture. Amplifying a polymorphic portion of the X chromosome-linked phosphoglycerate kinase (PGK) gene after selective digestion of the active X chromosome with a methylation-sensitive restriction enzyme gave results fully concordant with standard Southern blotting of DNA samples from normal (polyclonal) polymorphonuclear cells (PMN) as well as clonal PMN from patients with myelodysplastic syndrome and polycythemia vera (PCV). We have used this technique to demonstrate heterogeneity of lineage involvement in patients with PCV. The same clinical phenotype may arise from clonal proliferation of different hematopoietic progenitors.</jats:p>

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