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[An interlaboratory comparison study on the detection of RUNX1-RUNX1T1 fusion transcript levels and WT1 transcript

Y Z Qin1, L W Zhu2, S Lin3

  • 1Peking University People's Hospital, Peking University Institute of Hematology, National Clinical Research Center for Hematologic Disease, Beijing 100044, China.

Zhonghua Xue Ye Xue Za Zhi = Zhonghua Xueyexue Zazhi
|December 20, 2019
PubMed
Summary

This study assessed RUNX1-RUNX1T1 and WT1 transcript detection in China. While most labs showed high consistency, transcript levels varied significantly between laboratories for the same sample.

Keywords:
Fusion protein, RUNX1-RUNX1T1Interlaboratory comparisonReal-time quantitative PCRWT1

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Area of Science:

  • Molecular Diagnostics
  • Hematologic Malignancies
  • Interlaboratory Quality Assessment

Background:

  • Accurate detection of RUNX1-RUNX1T1 fusion transcripts and WT1 transcripts is crucial for diagnosing and monitoring acute myeloid leukemia.
  • Interlaboratory variability in quantitative PCR assays can impact clinical decision-making.

Purpose of the Study:

  • To evaluate the performance and consistency of RUNX1-RUNX1T1 and WT1 transcript level detection across different laboratories in China.
  • To identify potential sources of variability in real-time quantitative PCR assays for these transcripts.

Main Methods:

  • A comparison study was conducted using serially diluted bone marrow samples with known RUNX1-RUNX1T1 status.
  • Twenty laboratories performed real-time quantitative PCR to quantify RUNX1-RUNX1T1 and WT1 transcript levels relative to ABL.
  • Spearman correlation coefficients were calculated to assess agreement with a central reference laboratory (PKUPH).

Main Results:

  • False negative and positive rates for RUNX1-RUNX1T1 were low (0% and 5%, respectively).
  • Significant variability in reported transcript levels was observed, with ratios of highest to lowest measurements ranging from 5.3 to 13.7.
  • While most laboratories (85% for RUNX1-RUNX1T1, 62% for WT1) showed high correlation (≥0.98) with the reference, the relative performance varied between the two transcripts.

Conclusions:

  • Substantial interlaboratory variation exists in the quantification of RUNX1-RUNX1T1 and WT1 transcripts.
  • Most laboratories demonstrate good agreement with a reference standard, but consistency across different transcript targets is not guaranteed.
  • Standardization efforts are needed to improve the reliability of molecular diagnostic testing for hematologic malignancies.