Journal
JOURNAL OF MOLECULAR DIAGNOSTICS
Volume 19, Issue 3, Pages 437-444Publisher
ELSEVIER SCIENCE INC
DOI: 10.1016/j.jmoldx.2017.01.004
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Funding
- Savings Bank Foundation of Puglia
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Nested RT-PCR (nPCR) and real-time quantitative PCR (qPCR) are well-established methods for monitoring minimal residual disease (MRD) in acute promyelocytic Leukemia (APL). Despite their remarkable sensitivity and specificity, both methods have inherent limitations, such as qualitative MRD evaluation and relative quantification. Herein, we used droplet digital PCR (ddPCR) to monitor MRD in 21 APL patients and compared its performance with nPCR and qPCR. After assessing the Limit of detection (LOD) for each technique on serial dilutions of PML-RARA bcrl and bcr3 transcripts, a total of 48 follow-up samples were analyzed and the results compared. ddPCR showed good Linearity and efficiency and reached an LOD comparable or even superior to nPCR and qPCR. When tested on primary samples, ddPCR exhibited a sensitivity and specificity of >= 95% and >= 91% for bcrl and bcr3 transcripts and displayed a significant concordance with both techniques, particularly with nPCR. The peculiar advantage of ddPCR-based monitoring of MRD is represented by absolute quantification, which provides crucial information for the management of patients whose MRD fluctuates under the LOD of qPCR and is detectable, but not quantifiable, by nPCR. Ourfindings highlight ddPCR as a reliable complementary approach to monitor MRD in APL, and suggest its advantageous application, particularly for the molecular follow-up of patients at high risk of relapse.
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