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A certified plasmid reference material for the standardisation of BCR-ABL1 mRNA quantification by real-time
H White1, L Deprez2, P Corbisier2
11] National Genetics Reference Laboratory (Wessex), Salisbury District Hospital, Salisbury, UK [2] Faculty of Medicine, University of Southampton, Southampton, UK.
Leukemia
|July 19, 2014
Summary
Standardized BCR-ABL1 mRNA quantification is crucial for chronic myeloid leukaemia treatment monitoring. A new plasmid certified reference material (CRM) was developed to improve laboratory result comparability and accuracy.
Area of Science:
- Molecular Diagnostics
- Biotechnology
- Clinical Chemistry
Background:
- Serial quantification of BCR-ABL1 mRNA is vital for monitoring chronic myeloid leukaemia (CML) treatment efficacy.
- Significant inter-laboratory variability in BCR-ABL1 mRNA measurements hinders reliable therapeutic assessment.
- Standardization of molecular diagnostic assays is essential for accurate patient management.
Purpose of the Study:
- To develop and certify an internationally accepted plasmid reference material for BCR-ABL1 mRNA quantification.
- To enhance the comparability and accuracy of BCR-ABL1 mRNA measurements across different laboratories.
- To provide a standardized tool for validating diagnostic assays used in CML management.
Main Methods:
- Development of a plasmid (pIRMM0099) containing BCR-ABL1 (e14a2 fusion), BCR, and GUSB transcripts.
- Certification of six linearized plasmid solutions with precise copy number concentrations using digital PCR.
- Assessment of material heterogeneity and stability according to ISO Guide 34:2009 and ISO Guide 35:2006.
Main Results:
- Six plasmid certified reference materials (CRMs) with assigned copy number concentrations and uncertainties were produced.
- Suitability studies involving 63 laboratories demonstrated the CRMs' ability to standardize e14a2 BCR-ABL1 transcript measurements.
- The CRMs also aided in standardizing the quantification of control genes ABL1, BCR, and GUSB.
Conclusions:
- The developed set of six plasmid CRMs provides a robust tool for standardizing BCR-ABL1 mRNA quantification in CML diagnostics.
- Implementation of these CRMs can significantly reduce inter-laboratory variation, leading to more reliable therapeutic monitoring.
- These reference materials are globally available, promoting consistency in CML patient care worldwide.
