An Automated Cartridge-Based Microfluidic System for Real-Time Quantification of BCR::ABL1 Transcripts in Chronic Myeloid Leukemia: An Italian Experience

Alice Costanza Danzero; Enrico Marco Gottardi; Fabrizio Quarantelli; Ciro Del Prete; Alessandra Potenza; Claudia Venturi; Paola Berchialla; Francesca Guerrini; Clara Bono; Emanuela Ottaviani; Sara Galimberti; Carmen Fava; Barbara Izzo

doi:10.3390/ijms26188932

journal article Open Access Sep 13, 2025

An Automated Cartridge-Based Microfluidic System for Real-Time Quantification of BCR::ABL1 Transcripts in Chronic Myeloid Leukemia: An Italian Experience

Alice Costanza Danzero

Enrico Marco Gottardi Fabrizio Quarantelli

Ciro Del Prete Alessandra Potenza Claudia Venturi Paola Berchialla

Francesca Guerrini Clara Bono Emanuela Ottaviani Sara Galimberti Carmen Fava Barbara Izzo

International Journal of Molecular Sciences Vol. 26 No. 18 pp. 8932 · MDPI AG

View at Publisher Save 10.3390/ijms26188932

Abstract

Chronic myeloid leukemia (CML) is a clonal myeloproliferative disorder caused by the BCR::ABL1 fusion gene, resulting from a reciprocal translocation between chromosomes 22 and 9. Quantification of BCR::ABL1 transcript levels in peripheral blood by RT-qPCR represents the gold standard for molecular response (MR) monitoring, providing essential clinical information on treatment efficacy. Xpert® BCR-ABL Ultra is a fully automated in vitro diagnostic test that quantitatively detects e13a2 and e14a2 BCR::ABL1 transcripts using a single-use cartridge that integrates RNA extraction, cDNA synthesis, nested real-time PCR, and signal detection within a rapid, closed, and user-friendly system. In this study, we evaluated Xpert® BCR-ABL Ultra as an alternative to validated systems currently used by four highly specialized Italian laboratories affiliated with the Italian national laboratory network for CML. A total of 129 peripheral blood samples from CML patients at various disease stages, along with two external quality control materials, were analyzed. We assessed the test’s repeatability, specificity, and stability. Concordance of BCR::ABL1%IS values generated by the different methods was evaluated using EUTOS criteria and Bland–Altman analysis. Finally, MR value concordance was analyzed based on European LeukemiaNet recommendations or calculated using the formula 2 − log10(BCR::ABL1%IS). Xpert® BCR-ABL Ultra demonstrated high repeatability and stability. The BCR::ABL1%IS values obtained with this assay showed strong concordance with those generated by local reference methods, and MR classifications were consistent across platforms. These findings confirm the robustness, accuracy, and efficiency of the Xpert® BCR-ABL Ultra assay, supporting its use as a reliable alternative to currently validated systems for the routine clinical monitoring of CML patients.

Topics

No keywords indexed for this article. Browse by subject →

References

18

[1]

Lichty "Expression of p210 and p190 BCR-ABL due to alternative splicing in chronic myelogenous leukemia" Br. J. Haematol. (1998) 10.1046/j.1365-2141.1998.01033.x

[2]

Baccarani "The proportion of different BCR-ABL1 transcript types in chronic myeloid leukemia. An international overview" Leukemia (2019) 10.1038/s41375-018-0341-4

[3]

Apperley "2025 European LeukemiaNet recommendations for the management of chronic myeloid leukemia" Leukemia (2025) 10.1038/s41375-025-02664-w

[4]

Cross "European LeukemiaNet laboratory recommendations for the diagnosis and management of chronic myeloid leukemia" Leukemia (2023) 10.1038/s41375-023-02048-y

[5]

Chronic myeloid leukaemia: ESMO Clinical Practice Guidelines for diagnosis, treatment and follow-up

A. Hochhaus, S. Saussele, G. Rosti et al.

Annals of Oncology 2017 10.1093/annonc/mdx219

[6]

Dufresne "Quantitative assessment of the BCR-ABL transcript using the Cepheid Xpert BCR-ABL Monitor assay" Arch. Pathol. Lab. Med. (2007) 10.5858/2007-131-947-qaotbt

[7]

Jovanovski, A., Petiti, J., Giugliano, E., Gottardi, E.M., Saglio, G., Cilloni, D., and Fava, C. (2020). Standardization of BCR-ABL1 p210 Monitoring: From Nested to Digital PCR. Cancers, 12. 10.3390/cancers12113287

[8]

Dominy "Evaluation of Xpert® BCR-ABL Ultra for the confirmation of BCR-ABL1 international scale conversion factors for the molecular monitoring of chronic myeloid leukaemia" Int. J. Lab. Hematol. (2021) 10.1111/ijlh.13348

[9]

Baccarani "European LeukemiaNet recommendations for the management of chronic myeloid leukemia: 2013" Blood (2013) 10.1182/blood-2013-05-501569

[10]

Carstensen "Comparing methods of measurement: Extending the LoA by regression" Stat. Med. (2010) 10.1002/sim.3769

[11]

Hughes "Monitoring CML patients responding to treatment with tyrosine kinase inhibitors: Review and recommendations for harmonizing current methodology for detecting BCR-ABL transcripts and kinase domain mutations and for expressing results" Blood (2006) 10.1182/blood-2006-01-0092

[12]

Hughes "Moving treatment-free remission into mainstream clinical practice in CML" Blood (2016) 10.1182/blood-2016-01-694265

[13]

Lippi "Blood sample quality" Diagnosis (2019) 10.1515/dx-2018-0018

[14]

Cepheid (2025, August 16). Xpert BCR-ABL Ultra Package Insert. Sunnyvale. (CE-IVD) 302-0742, January 2021. Available online: https://web-support.cepheid.com/Package%20Insert%20Files/Xpert%20BCR-ABL%20Ultra%20Assay-ENGLISH%20PI%20302-0738%20Rev.%20C_1.pdf.

[15]

Macintyre "Standardized RT-PCR analysis of fusion gene transcripts from chromosome aberrations in acute leukemia for detection of minimal residual disease. Report of the BIOMED-1 Concerted Action: Investigation of minimal residual disease in acute leukemia" Leukemia (1999) 10.1038/sj.leu.2401592

[16]

Cross "Harmonization of molecular monitoring of CML therapy in Europe" Leukemia (2009) 10.1038/leu.2009.168

[17]

Cross "Standardized definitions of molecular response in chronic myeloid leukemia" Leukemia (2012) 10.1038/leu.2012.104

[18]

White "Standardization of molecular monitoring of CML: Results and recommendations from the European treatment and outcome study" Leukemia (2022) 10.1038/s41375-022-01607-z

Metrics

1

Citations

18

References

Details

Published: Sep 13, 2025
Vol/Issue: 26(18)
Pages: 8932
License: View

Authors

A

Alice Costanza Danzero

Department of Clinical and Biological Sciences, Turin University, 10043 Orbassano, Italy

E

Enrico Marco Gottardi

Department of Clinical and Biological Sciences, Turin University, 10043 Orbassano, Italy

F

Fabrizio Quarantelli

Oncologic Hematology and Cytogenetics Laboratory, CEINGE Advanced Biotechnology “Franco Salvatore”, 80131 Naples, Italy

C

Ciro Del Prete

Oncologic Hematology and Cytogenetics Laboratory, CEINGE Advanced Biotechnology “Franco Salvatore”, 80131 Naples, Italy

A

Alessandra Potenza

Oncologic Hematology and Cytogenetics Laboratory, CEINGE Advanced Biotechnology “Franco Salvatore”, 80131 Naples, Italy

C

Claudia Venturi

“Seràgnoli” Institute of Hematology, IRCCS University Hospital of Bologna, 40138 Bologna, Italy

P

Paola Berchialla

Department of Clinical and Biological Sciences, Turin University, 10043 Orbassano, Italy

F

Francesca Guerrini

Department of Clinical and Experimental Medicine, Hematology, University of Pisa, 56126 Pisa, Italy

C

Clara Bono

Department of Clinical and Experimental Medicine, Hematology, University of Pisa, 56126 Pisa, Italy

E

Emanuela Ottaviani

“Seràgnoli” Institute of Hematology, IRCCS University Hospital of Bologna, 40138 Bologna, Italy

S

Sara Galimberti

Department of Clinical and Experimental Medicine, Hematology, University of Pisa, 56126 Pisa, Italy

C

Carmen Fava

Department of Clinical and Biological Sciences, Turin University, 10043 Orbassano, Italy

B

Barbara Izzo

Department of Molecular Medicine and Medical Biotechnology, University of Naples “Federico II” and CEINGE Advanced Biotechnology “Franco Salvatore”, 80131 Naples, Italy

Funding

Universities of Naples, Turin, Bologna, and Pisa

Cepheid

Cite This Article

Alice Costanza Danzero, Enrico Marco Gottardi, Fabrizio Quarantelli, et al. (2025). An Automated Cartridge-Based Microfluidic System for Real-Time Quantification of BCR::ABL1 Transcripts in Chronic Myeloid Leukemia: An Italian Experience. International Journal of Molecular Sciences, 26(18), 8932. https://doi.org/10.3390/ijms26188932

An Automated Cartridge-Based Microfluidic System for Real-Time Quantification of BCR::ABL1 Transcripts in Chronic Myeloid Leukemia: An Italian Experience

You May Also Like