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Variant-specific BCR::ABL1 quantification discrepancy in chronic myeloid leukemia.

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|June 6, 2024
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Summary

Accurate quantification of BCR::ABL1 fusion gene transcripts is crucial for chronic myeloid leukemia (CML) management. Using transcript-specific standard curves or absolute quantification methods reduces inter-center variability in BCR::ABL1 measurements.

Keywords:
BCR‐ABLCMLminimal residual disease

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Area of Science:

  • Molecular diagnostics
  • Oncology
  • Clinical chemistry

Background:

  • Accurate quantification of the BCR::ABL1 fusion gene in whole blood is vital for managing chronic myeloid leukemia (CML).
  • BCR::ABL1 fusion protein size varies based on gene breakpoints, with most CML patients having the p210 BCR::ABL1 fusion gene.
  • The p210 BCR::ABL1 fusion gene typically arises from e14a2 (b3a2) or e13a2 (b2a2) mRNA transcript junctions.

Purpose of the Study:

  • To assess inter-center variability in BCR::ABL1 quantification using quantitative polymerase chain reaction (qPCR).
  • To evaluate the impact of transcript-specific standard curves and digital droplet PCR (ddPCR) on reducing quantification differences.
  • To highlight the clinical implications of quantification discrepancies in CML patient monitoring.

Main Methods:

  • Analyzed 25 CML samples across two ISO15189-accredited centers using a standardized Europe Against Cancer-based qPCR protocol.
  • Performed reanalysis with transcript-specific standard curves and digital droplet PCR (ddPCR).
  • Compared qPCR results from both centers with ddPCR, an absolute quantification method.

Main Results:

  • Significant inter-center differences (up to 1 log) were observed for the e13a2 transcript variant, but not for e14a2 transcripts.
  • Reanalysis using transcript-specific standard curves eliminated these inter-center differences.
  • Transcript-specific qPCR results showed significant association with ddPCR, particularly at lower quantification ranges.

Conclusions:

  • Inter-center differences in transcript-specific BCR::ABL1 quantification can impact CML patient management.
  • Employing transcript-specific standard curves or absolute quantification methods significantly minimizes these discrepancies.
  • Careful interpretation of quantification differences is essential, especially for CML patients transferring between diagnostic centers.