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A new DPYD genotyping assay for improving the safety of 5-fluorouracil therapy
Johanna Sistonen1, Chingying Smith, Yung-Kang Fu
1Institute of Clinical Chemistry, Inselspital, Bern University Hospital, and University of Bern, Bern, Switzerland. johanna.sistonen@insel.ch
Background:
Chemotherapeutic use of 5-fluorouracil (5FU) is compromised by 10-20% of patients developing severe toxicity. Recently described genetic variation in dihydropyrimidine dehydrogenase (DPYD) has been shown to be a major predictor of 5FU toxicity. Here, we describe a new genotyping assay for routine clinical use that covers all the major DPYD risk variants.
Methods:
Genomic regions targeting DPYD risk variants (c.1129-5923C>G, c.1679T>G/A, c.1905+1G>A, c.2846A>T) and additional markers (c.234-123G>C, c.496A>G, c.775A>G) were amplified in a multiplex PCR reaction. The subsequent steps including allele-specific primer extension, hybridization of the primers to a microarray, scanning of the array, and data analysis were automated within the INFINITI® Analyzer (AutoGenomics). The assay was validated by analyzing 107 blood samples obtained from patients previously re-sequenced for the DPYD.
Results:
The genotypes obtained with the developed assay were 100% concordant with the re-sequencing. The procedure is suitable for routine clinical use since the results are obtained within one day. For heterozygous risk variant carriers (~7% of Europeans), the treatment can be adjusted by 5FU dose reduction, whereas carriers of two risk alleles should be treated with an alternative therapy.
Conclusions:
The developed assay provides a novel tool to improve the safety of commonly used 5FU-based chemotherapies.
Insights
A new DPYD genotyping assay identifies patients at risk for severe 5-fluorouracil (5FU) toxicity. This test enables personalized chemotherapy, improving patient safety by adjusting 5FU dosage or using alternative treatments.
Area of Science:
- Pharmacogenomics
- Clinical Chemistry
- Molecular Diagnostics
Background:
- Severe toxicity affects 10-20% of patients undergoing 5-fluorouracil (5FU) chemotherapy.
- Genetic variations in dihydropyrimidine dehydrogenase (DPYD) are key predictors of 5FU toxicity.
- A need exists for a reliable assay to identify DPYD risk variants for clinical application.
Purpose of the Study:
- To develop and validate a novel, automated genotyping assay for routine clinical detection of major DPYD risk variants.
- To facilitate personalized 5FU chemotherapy by identifying patients susceptible to severe adverse events.
Main Methods:
- Multiplex PCR amplification of DPYD genomic regions harboring key risk variants.
- Automated allele-specific primer extension, microarray hybridization, and data analysis using the INFINITI® Analyzer.
- Validation of the assay against patient samples previously analyzed by re-sequencing.
Main Results:
- The developed assay demonstrated 100% concordance with established re-sequencing methods.
- Results are available within one day, making the assay suitable for routine clinical use.
- Identified heterozygous carriers (~7% of Europeans) can benefit from 5FU dose adjustment, while homozygous carriers require alternative therapies.
Conclusions:
- The new DPYD genotyping assay is a valuable tool for enhancing the safety of 5FU-based chemotherapies.
- Clinical implementation of this assay can significantly reduce 5FU-related toxicity.
- Personalized medicine approaches guided by DPYD genotyping improve patient outcomes in cancer treatment.
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