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  1. Home
  3. Establishment Of Potential Reference Measurement Procedure And Reference Materials For Eml4-alk Fusion Variants Measurement.
  1. Home
  3. Establishment Of Potential Reference Measurement Procedure And Reference Materials For Eml4-alk Fusion Variants Measurement.

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Establishment of potential reference measurement procedure and reference materials for EML4-ALK fusion variants

Yi Yang1,2, Yu Zhang1, Shujun Zhou2

  • 1Center for Advanced Measurement Science, National Institute of Metrology, Beijing, 100029, China.

Scientific Reports
|October 19, 2024

View abstract on PubMed

Summary
This summary is machine-generated.

Establishing traceability for Echinoderm microtubule-associated protein 4 (EML4)-anaplastic lymphoma kinase (ALK) fusion gene detection is crucial for personalized cancer therapy. This study developed a reference measurement procedure and reference materials using RT-dPCR to ensure consistent and comparable clinical test results.

Keywords:
EML4-ALKNon-small-cell lung cancerReference materials (RMs)Reference measurement procedure (RMP)Reverse transcription dPCR (RT-dPCR)

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Area of Science:

  • Molecular Diagnostics
  • Biotechnology
  • Clinical Chemistry

Background:

  • Accurate detection of Echinoderm microtubule-associated protein 4 (EML4)-anaplastic lymphoma kinase (ALK) fusion genes is vital for targeted cancer therapy.
  • Ensuring consistency and comparability of EML4-ALK detection results across different laboratories is a clinical challenge.
  • Traceability in molecular diagnostics relies on traceable reference materials and reference measurement procedures.

Purpose of the Study:

  • To establish a potential reference measurement procedure (RMP) for quantifying EML4-ALK fusion variants (V1 and V3) and reference types (ALK-ref).
  • To develop EML4-ALK fusion gene reference materials (RMs) for establishing measurement traceability.
  • To validate the accuracy and reproducibility of the developed RMP and RMs in inter-laboratory studies.

Main Methods:

  • Development of a potential reference measurement procedure (RMP) based on reverse transcription digital PCR (RT-dPCR).
  • Creation of EML4-ALK fusion gene reference materials (RMs) including wild type and V1/V3 mutant types.
  • Conducting inter-laboratory studies to assess the performance of the RMP and RMs.

Main Results:

  • The potential RMP demonstrated high specificity, good inter-laboratory reproducibility (CV < 7.3%), and excellent linearity (R² ≥ 0.99).
  • The limit of detection (LoD) for V1 and V3 variants was 2 copies/reaction, and for ALK-ref was 1 copy/reaction.
  • Inter-laboratory studies confirmed consistent copy concentrations and ratios, indicating successful establishment of traceability.

Conclusions:

  • The developed RT-dPCR based potential RMP and EML4-ALK RMs are suitable for characterizing reference materials.
  • The established RMP and RMs provide a foundation for traceable EML4-ALK fusion measurements.
  • This work significantly improves the comparability and consistency of EML4-ALK detection in clinical practice, supporting personalized tumor treatment.