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Analytical and Bioanalytical Chemistry

, Volume 411, Issue 4, pp 895–903 | Cite as

Droplet digital PCR for quantification of PML-RARα in acute promyelocytic leukemia: a comprehensive comparison with real-time PCR

  • Dandan Yuan
  • Ming Cui
  • Shuping Yu
  • Huimin WangEmail author
  • Rongrong JingEmail author
Research Paper

Abstract

Real-time quantitative PCR (qPCR) has been widely implemented for molecular testing, but there are still some inherent limitations that hamper its usefulness. Droplet digital PCR (ddPCR), which can provide direct, standards-free quantification, has recently received increasing attention. In our study, a comprehensive comparison of ddPCR with qPCR in relation to the quantification of PML-RARα was performed to evaluate the diagnostic potential of ddPCR. Results showed that ddPCR displayed significant concordance with qPCR in the detection of PML-RARα in clinical samples, but showed advantages over qPCR in terms of precision, limit of detection (LOD), and other basic performance parameters. A study of the feasibility of duplexing also indicated that ddPCR could simultaneously quantify the target PML-RARα and the clinical common reference gene ABL in a reaction, in contrast to qPCR. Moreover, ddPCR was more tolerant than qPCR of inhibition, and was shown to be able to quantify inhibition-prone samples. Another advantage of using ddPCR in clinical applications is that it will yield accurate results for patients with PML-RARα levels that fluctuate around the LOD of qPCR. Therefore, ddPCR is considered to have the potential to become a reliable alternative technique for quantifying PML-RARα.

Graphical abstract

Keywords

ddPCR qPCR PML-RARα quantification Comparison of methods 

Notes

Acknowledgements

This study was supported by the National Natural Science Foundation (grant numbers 81201349, 81000775); Young Medical Key Talents in Jiangsu Province (grant numbers QNRC 2016686, 2016687); Frontier and Key Technical Innovation Projects of Nantong (grant number MS22015049); and the Nantong Science and Technology Plan Project (MS12017008-3).

Compliance with ethical standards

The authors declare that all individual participants from whom the blood samples were obtained gave their informed consent, and that the studies were approved by the ethics committee of the Affiliated Hospital of Nantong University and were performed in accordance with ethical standards.

Conflicts of interest

The authors declare no conflict of interest.

Supplementary material

216_2018_1508_MOESM1_ESM.pdf (323 kb)
ESM 1 (PDF 322 kb)

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Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Department of Laboratory MedicineAffiliated Hospital of Nantong UniversityNantongChina

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