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

Cancer Chemother Pharmacol. 2015 May;75(5):1095-8. doi: 10.1007/s00280-015-2729-4. Epub 2015 Mar 25.

Abstract

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.

Publication types

  • Research Support, Non-U.S. Gov't
  • Validation Study

MeSH terms

  • Alkalies / chemistry
  • Colorectal Neoplasms / blood*
  • Colorectal Neoplasms / drug therapy
  • Colorectal Neoplasms / genetics*
  • DNA, Neoplasm / blood
  • DNA, Neoplasm / genetics
  • DNA, Neoplasm / isolation & purification*
  • Dihydrouracil Dehydrogenase (NADP) / genetics
  • Fluorouracil / toxicity
  • Genotyping Techniques
  • Humans
  • Mutation
  • Pharmacogenetics
  • Prospective Studies
  • Reproducibility of Results

Substances

  • Alkalies
  • DNA, Neoplasm
  • Dihydrouracil Dehydrogenase (NADP)
  • Fluorouracil