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Establishment of genomic RNA reference materials for BCR-ABL1 P210 measurement.

Author information

Affiliations

  • 1. Center for Advanced Measurement Science, National Institute of Metrology, Beijing, 100029, China.
      Authors
    • Yang Y 1, 2
    • Wang X 1
    • Niu C 1
    • Dong L 1
    (4 authors)
  • 2. Shenzhen Institute for Technology Innovation, National Institute of Metrology, Shenzhen, 518107, China.
      Authors
    • Yang Y 1, 2
    • Zhou S 2
    (2 authors)
  • 3. National Research Institute for Family Planning, Beijing, 100081, China.
      Authors
    • Gao H 3
    • Jin X 3
    (2 authors)
  • 4. Nanjing Institute of Measurement and Testing Technology, Nanjing, 210049, Jiangsu, China.
      Authors
    • Wang S 4
    (1 author)
  • 5. Institute of Analysis and Testing, Beijing Academy of Science and Technology (Beijing Center for Physical and Chemical Analysis), Beijing, 100094, China.
      Authors
    • Du M 5
    • Cheng X 5
    (2 authors)
    • Show all (6)

Analytical and Bioanalytical Chemistry, 09 Sep 2024, 416(26):5733-5742
https://doi.org/10.1007/s00216-024-05492-6 PMID: 39251426 

Abstract 

Quantitation of BCR-ABL1 with the quantitative reverse transcriptase polymerase chain reaction (RT-PCR) is very important in monitoring chronic myeloid leukemia (CML), which relies on an RNA reference material. A genomic RNA reference material (RM) containing the BCR-ABL1 P210 fusion mutation was developed, and an absolute quantitative method based on one-step reverse transcription digital PCR (RT-dPCR) was established for characterizing the RM. The proposed dPCR method demonstrates high accuracy and excellent analytical sensitivity, as shown by the linear relationship (0.94 < slope < 1.04, R2≧0.99) between the measured and nominal values of b2a2, b3a2, and ABL1-ref within the dynamic range (104-101 copies/reaction). Homogeneity and stability assessment based on dPCR indicated that the RM was homogeneous and stable for 24 months at -80 °C. The RM was used to evaluate inter-laboratory reproducibility in eight different laboratories, demonstrating that participating laboratories could consistently produce copy concentrations of b3a2 and ABL1-ref, as well as the BCR-ABL1/ABL1 ratio (CV < 2.0%). This work suggests that the RM can be employed in establishing metrological traceability for detecting mutations in the BCR-ABL1 fusion gene, as well as in quality control for testing laboratories.

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Read article at publisher's site: https://doi.org/10.1007/s00216-024-05492-6

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    Funding 

    Funders who supported this work.

    National Key Research and Development Program of China (1)