Sensitive detection and quantification of minimal residual disease in chronic myeloid leukaemia using nested quantitative PCR for BCR-ABL DNA

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dc.contributor.authorBartley, P.
dc.contributor.authorRoss, D.
dc.contributor.authorLatham, S.
dc.contributor.authorMartin-Harris, M.
dc.contributor.authorBudgen, B.
dc.contributor.authorWilczek, V.
dc.contributor.authorBranford, S.
dc.contributor.authorHughes, T.
dc.contributor.authorMorley, A.
dc.date.issued2010
dc.description.abstractIncreasing numbers of patients with chronic myeloid leukaemia (CML) treated with tyrosine kinase inhibitors achieve undetectable levels of BCR-ABL mRNA using sensitive quantitative real-time reverse transcriptase PCR (RT-qPCR) methods and a method to measure minimal residual disease (MRD) in patients with low levels could be of value. Following isolation and sequencing of the patient-specific BCR-ABL breakpoint, a DNA-based nested qPCR assay was established, and MRD was measured by this method and one-round RT-qPCR in 38 samples from 24 patients with CML. Mixing experiments using patient DNA in normal DNA indicated that DNA qPCR could detect BCR-ABL sequences at a limit of approximately 10⁻⁶. In 22 samples in which MRD was detectable by both methods, comparison of the results of DNA qPCR with the results obtained on the same sample by RT-qPCR showed good correlation. In another 16 samples, BCR-ABL mRNA was not detectable by RT-qPCR. In 8 of the 16 samples, BCR-ABL DNA was detected at levels ranging from 1.1 × 10⁻⁵ up to 2.8 × 10⁻⁴ and in the remaining eight samples BCR-ABL was not detected by either method. In one patient, who had stopped imatinib, an almost 1000-fold rise in MRD, to 5.2 × 10⁻⁴ was observed in sequential samples. Nested DNA qPCR was more sensitive than one-round RT-qPCR and could be used for the monitoring of patients with CML with very low levels of MRD.
dc.description.statementofresponsibilityP. A. Bartley, D. M. Ross, S. Latham, M. H. Martin-Harris, B. Budgen, V. Wilczek, S. Branford, T. P. Hughes, A. A. Morley
dc.identifier.citationInternational Journal of Laboratory Hematology, 2010; 32(6 Part 1):E222-E228
dc.identifier.doi10.1111/j.1751-553X.2010.01236.x
dc.identifier.issn1751-5521
dc.identifier.issn1751-553X
dc.identifier.orcidRoss, D. [0000-0001-7171-2935]
dc.identifier.orcidBranford, S. [0000-0002-1964-3626] [0000-0002-5095-7981]
dc.identifier.orcidHughes, T. [0000-0002-0910-3730] [0000-0002-7990-4509]
dc.identifier.urihttp://hdl.handle.net/2440/63823
dc.language.isoen
dc.publisherBlackwell Publishing Ltd
dc.rights© 2010 Blackwell Publishing Ltd.
dc.source.urihttps://doi.org/10.1111/j.1751-553x.2010.01236.x
dc.subjectMinimal residual disease
dc.subjectchronic myeloid leukaemia
dc.subjectBCR-ABL
dc.subjectPCR
dc.subjectmonitoring
dc.titleSensitive detection and quantification of minimal residual disease in chronic myeloid leukaemia using nested quantitative PCR for BCR-ABL DNA
dc.typeJournal article
pubs.publication-statusPublished

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