Rapid isolation of translocation breakpoints in chronic myeloid and acute promyelocytic leukaemia: Research paper

Paul Bartley, Michael Martin-Harris, Bradley Budgen, David Ross, Alexander Morley

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    17 Citations (Scopus)

    Abstract

    Isolation and sequencing of the translocation breakpoint in chronic myeloid leukaemia-(CML) and acute promyelocytic leukaemia (APML) may help to elucidate the mechanism of translocation and provide a molecular marker for monitoring of minimal residual disease. Amplification across the translocation breakpoint was performed in samples from 91 patients with CML and 15 patients with APML using single-tube multiplex polymerase chain reaction (PCR) involving 308 primers for CML and 40 primers for APML. Nonspecific amplification was removed by a modification of PCR, termed sequential hybrid primer PCR (SHP-PCR), which involved two sequential rounds of PCR, each of which used a low concentration of a specially designed hybrid primer. The resultant amplified material was sequenced. The method as finally developed was simple quick and robust. The translocation breakpoint was successfully isolated and sequenced in all 106 samples. The strategy of highly multiplexed PCR followed by SHP-PCR is thus an effective method for isolating the translocation breakpoint in CML and APML. It may also be applicable to other haematological disorders characterised by translocation, deletion or inversion.

    Original languageEnglish
    Pages (from-to)231-236
    Number of pages6
    JournalBritish Journal of Haematology
    Volume149
    Issue number2
    DOIs
    Publication statusPublished - Apr 2010

    Keywords

    • Acute promyelocytic leukaemia
    • BCR-ABL1
    • Chronic myeloid leukaemia
    • PML-RARA
    • Polymerase chain reaction
    • Translocations

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