Fast CYP2E1 genotyping using automated fluorescent detection

Med Lav. 2006 Nov-Dec;97(6):799-804.

Abstract

Background: Among polymorphic genes coding for xenobiotic-metabolizing enzymes, the ethanol-inducible CYP2E1 gene (1667 bp) is known to play a major role in the metabolism ofseveral chemicals.

Objectives: In order to apply large-scale genotyping, we explored the use of a Single Nucleotide Primer Extension (SNuPE) assay coupled with automated fluorescent detection to assess the presence of low-frequency CYP2E1*5B (c2) allele.

Methods: a classic PCR-RFLP (Polymerase Chain Reaction-Restriction Fragment Length Polymorphism) method specific for polymorphic5'-flanking region of CYP2E1 gene was tested in conjunction with a newly developed accelerated SNuPE assay.

Results: compared to the classic PCR-RFLP method, the accelerated SNuPE assay proved to be both sensitive and specific for fast CYP2E1 genotyping.

Conclusions: automated fluorescent methods as SNuPE assay are usefulfor public health perspectives, allowing rapid genotyping of metabolic genes in large population studies in clinical or epidemiological settings.

Publication types

  • Comparative Study

MeSH terms

  • Alleles
  • Cytochrome P-450 CYP2E1 / genetics*
  • DNA Primers*
  • Electrophoresis, Agar Gel
  • Female
  • Fluorescence*
  • Genotype*
  • Humans
  • Male
  • Polymerase Chain Reaction
  • Polymorphism, Restriction Fragment Length
  • Sensitivity and Specificity
  • Software

Substances

  • DNA Primers
  • Cytochrome P-450 CYP2E1