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https://hdl.handle.net/2440/63823
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Type: | Journal article |
Title: | Sensitive detection and quantification of minimal residual disease in chronic myeloid leukaemia using nested quantitative PCR for BCR-ABL DNA |
Author: | Bartley, P. Ross, D. Latham, S. Martin-Harris, M. Budgen, B. Wilczek, V. Branford, S. Hughes, T. Morley, A. |
Citation: | International Journal of Laboratory Hematology, 2010; 32(6 Part 1):E222-E228 |
Publisher: | Blackwell Publishing Ltd |
Issue Date: | 2010 |
ISSN: | 1751-5521 1751-553X |
Statement of Responsibility: | P. A. Bartley, D. M. Ross, S. Latham, M. H. Martin-Harris, B. Budgen, V. Wilczek, S. Branford, T. P. Hughes, A. A. Morley |
Abstract: | Increasing 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. |
Keywords: | Minimal residual disease chronic myeloid leukaemia BCR-ABL PCR monitoring |
Rights: | © 2010 Blackwell Publishing Ltd. |
DOI: | 10.1111/j.1751-553X.2010.01236.x |
Published version: | http://dx.doi.org/10.1111/j.1751-553x.2010.01236.x |
Appears in Collections: | Aurora harvest 5 Medicine publications |
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