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Comparison of Droplet Digital PCR versus qPCR Measurements on the International Scale for the Molecular Monitoring of Chronic Myeloid Leukemia Patients

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Abstract

Background

BCR-ABL1/ABL1 p210 measurement by quantitative polymerase chain reaction (qPCR) is used worldwide to monitor the molecular response in chronic myeloid leukemia (CML) patients. Droplet digital polymerase chain reaction (ddPCR) seems to show a greater sensitivity than qPCR, probably due to the high number of replicates analyzed in ddPCR for the comparison. Additionally, in a recently published comparison, ddPCR measurements were not adequately transformed into International Scale (IS).

Method

We have analyzed 50 CML patients and ten non-CML donors in parallel by qPCR and ddPCR. To the best of our knowledge, this is the first study comparing both techniques under similar conditions, with BCR-ABL1/ABL1 measurements performed via both techniques transformed into IS.

Results

Qualitative and quantitative comparisons showed excellent results. The qualitative correlation showed a Kappa index of 0.94 (95% confidence interval [CI] 0.90–0.98) (P < 0.001). In the quantitative comparison, the absolute intra-class correlation coefficient was 0.868 (95% CI 0.734–0.937; P < 0.001), and Lin’s concordance correlation coefficient was 0.863. The Passing-Bablock test indicated a slight proportional difference between qPCR and ddPCR. A quantitative and qualitative subanalysis including 40 patients with a molecular response of 3.0 or deeper showed similar results in every test. In addition, the proportional difference in the Passing-Bablock test disappeared. There were no differences in the sensitivity for BCR-ABL1 detection between qPCR and ddPCR (McNemar test, P = 0.5).

Conclusions

In conclusion, our results show very good quantitative and qualitative correlations between BCR-ABL1/ABL1 p210 results obtained by qPCR and by ddPCR and confirm previous scarce data regarding the lack of an increase in sensitivity of ddPCR over qPCR in this setting.

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Authors and Affiliations

  1. Experimental Hematology, Instituto de Investigación Sanitaria Fundación Jiménez Díaz, Universidad Autónoma de Madrid, Avenida de los Reyes Católicos 2, 28040, Madrid, Spain

    Alicia Arenas Cortés, Juana Serrano-López, Daniel Lainez-González, Tamara Castaño, José Luis Lopez-Lorenzo, Mireia Atance, Rocío Nieves Salgado Sánchez, Carlos Blas Lopez, Pilar Llamas Sillero & Juan Manuel Alonso-Dominguez

  2. New Therapy Group, Instituto de Investigación Sanitaria Fundación Jiménez Díaz, Universidad Autónoma de Madrid, Madrid, Spain

    Susana Olmedillas & Mariano García Arranz

  3. Facultad de Farmacia, Universidad Complutense de Madrid, Madrid, Spain

    Juana Serrano-López

  4. Hematology Department, Fundación Jiménez Díaz University Hospital, Madrid, Spain

    Tamara Castaño, José Luis Lopez-Lorenzo, Amanda García, Mireia Atance, Rocío Nieves Salgado Sánchez, Carlos Blas Lopez, Pilar Llamas Sillero & Juan Manuel Alonso-Dominguez

  5. Hematology Department, Doce de Octubre University Hospital, Madrid, Spain

    Rodrigo Iñiguez

  6. Surgical Department, Medicine School, Universidad Autónoma de Madrid, Madrid, Spain

    Mariano García Arranz

Authors
  1. Alicia Arenas Cortés
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  2. Susana Olmedillas
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  3. Juana Serrano-López
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  4. Daniel Lainez-González
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  5. Tamara Castaño
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  6. Rodrigo Iñiguez
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  7. José Luis Lopez-Lorenzo
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  8. Amanda García
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  9. Mireia Atance
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  10. Rocío Nieves Salgado Sánchez
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  11. Carlos Blas Lopez
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  12. Mariano García Arranz
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  13. Pilar Llamas Sillero
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  14. Juan Manuel Alonso-Dominguez
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Corresponding author

Correspondence to Juan Manuel Alonso-Dominguez.

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Funding

This study was funded by Incyte S.L.

Conflicts of interest

Juan Manuel Alonso-Dominguez received research funding from Incyte Corporation, Pfizer International and Celgene International. AAR, SO, JSL, DLG, TC, RI, JLLL, AG, MA, RNSS, CBL, MGA, and PLS have no conflicts to declare.

Ethics approval

The protocol was approved by the Ethics Committee for Clinical Research of the institution (FJD-HEMPCR-16-01). Informed consent was not required according to the Ethics Committee for Clinical Research of the institution. Samples were collected in accordance with the Declaration of Helsinki.

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Cortés, A.A., Olmedillas, S., Serrano-López, J. et al. Comparison of Droplet Digital PCR versus qPCR Measurements on the International Scale for the Molecular Monitoring of Chronic Myeloid Leukemia Patients. Mol Diagn Ther 24, 593–600 (2020). https://doi.org/10.1007/s40291-020-00485-4

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  • DOI: https://doi.org/10.1007/s40291-020-00485-4

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