Validation of a fast and low-cost alkaline lysis method for gDNA extraction in a pharmacogenetic context

Purpose Translation of pharmacogenetic findings from the research laboratory to the clinical practice demands simple and efficient procedures. In this sense, we evaluated the suitability of a modified protocol for genomic DNA extraction based on alkaline lysis of cells. Methods Dried blood samples w...

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Published in:Cancer chemotherapy and pharmacology 2015-05, Vol.75 (5), p.1095-1098
Main Authors: Ramos-Díaz, Ruth, Gutiérrez-Nicolás, Fernando, Nazco-Casariego, Gloria J, González-Perera, Itamar, Pérez-Pérez, José A
Format: Article
Language:English
Subjects:
Alkalies - chemistry
Cancer Research
Colorectal Neoplasms - blood
Colorectal Neoplasms - drug therapy
Colorectal Neoplasms - genetics
Dihydrouracil Dehydrogenase (NADP) - genetics
DNA, Neoplasm - blood
DNA, Neoplasm - genetics
DNA, Neoplasm - isolation & purification
Fluorouracil - toxicity
Genotyping Techniques
Humans
Medicine
Medicine & Public Health
Mutation
Oncology
Pharmacogenetics
Pharmacology/Toxicology
Prospective Studies
Reproducibility of Results
Short Communication
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Summary:Purpose Translation of pharmacogenetic findings from the research laboratory to the clinical practice demands simple and efficient procedures. In this sense, we evaluated the suitability of a modified protocol for genomic DNA extraction based on alkaline lysis of cells. Methods Dried blood samples were obtained from 48 patients diagnosed with colorectal cancer. A total of 11 mutations in the dihydropyrimidine dehydrogenase gene and related to 5-fluorouracil toxicity were searched by amplicon sequencing and real-time PCR with fluorescent probes. Results Genomic DNA extracted with the alkaline lysis method, both from dried blood samples and buccal swabs, fulfilled the quality requirements of the two genotyping methods assayed, which yielded 100 % concordant results for 11 genetic variants with relevance to cancer chemotherapy. Conclusions The assessed protocol has shown to be a very fast and economical approach to perform genetic analyses in the clinical laboratory for pharmacological purposes.
ISSN:0344-5704
1432-0843
DOI:10.1007/s00280-015-2729-4