Journal
EUROPEAN JOURNAL OF HAEMATOLOGY
Volume 101, Issue 3, Pages 291-296Publisher
WILEY
DOI: 10.1111/ejh.13084
Keywords
BCR; ABL(P210); CML; droplet digital PCR; minimal residual disease; RT-qPCR
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Funding
- Shanxi Scholarship Council of China [2015-Key 5]
- Natural Science Foundation of China [81670126, 81400139]
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ObjectiveThis study intended to establish a droplet digital PCR (dd-PCR) for monitoring minimal residual disease (MRD) in patients with BCR/ABL (P210)-positive chronic myeloid leukemia (CML), thereby achieving deep-level monitoring of tumor load and determining the efficacy for guided clinically individualized treatment. MethodsUsing dd-PCR and RT-qPCR, two cell suspensions were obtained from K562 cells and normal peripheral blood mononuclear cells by gradient dilution and were measured at the cellular level. At peripheral blood (PB) level, 61 cases with CML-chronic phase (CML-CP) were obtained after tyrosine kinase inhibitor (TKI) treatment and regular follow-ups. By RT-qPCR, BCR/ABL (P210) fusion gene was undetectable in PB after three successive analyses, which were performed once every 3months. At the same time, dd-PCR was performed simultaneously with the last equal amount of cDNA. Ten CML patients with MR4.5 were followed up by the two methods. ResultsAt the cellular level, consistency of results of dd-PCR and RT-qPCR reached R(2)0.99, with conversion equation of Y=33.148X(1.222) (Y: dd-PCR results; X: RT-qPCR results). In the dd-PCR test, 11 of the 61 patients with CML (18.03%) tested positive and showed statistically significant difference (P<.01). In the follow-up of 10 CML patients who were in MR4.5. All patients were loss of MR4.0, and 4 were tested positive by dd-PCR 3months earlier than by RT-qPCR. ConclusionIn contrast with RT-qPCR, dd-PCR is more sensitive, thus enabling accurate conversion of dd-PCR results into internationally standard RT-qPCR results by conversion equation, to achieve a deeper molecular biology-based stratification of BCR/ABL(P210) MRD. It has some reference value to monitor disease progression in clinic.
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