Concordance between the deduced acetylation status generated by high-speed: real-time PCR based NAT2 genotyping of seven single nucleotide polymorphisms and human NAT2 phenotypes determined by a caffeine assay

Clin Chim Acta. 2007 Feb;376(1-2):240-3. doi: 10.1016/j.cca.2006.08.010. Epub 2006 Aug 14.

Abstract

Background: The utility of typing single nucleotide polymorphisms (SNPs) for the determination of the N-acetyltransferase 2 (NAT2) acetylation status is a matter of debate.

Aims of the study: Evaluation of the concordance between deduced genotype results of seven human NAT2 SNPs generated by Real-time PCR analysis and human NAT2 phenotypes.

Methods: NAT2 phenotypes of 38 Caucasian workers were determined using a suitable caffeine test method. Genomic DNA aliquots were used for the determination of seven human NAT2-specific SNPs (G191A, C282T, T341C, C481T, G590A, A803G, G857A).

Results and conclusions: Phenotypic results based on the molar ratio of 5-acetylamino-6-formylamino-3-methyluracil (AFMU)/(AFMU+1-methlyuric acid (1U)+1-methylxanthine (1X)) calculated from excreted caffeine metabolite levels in urine samples with 0.3 as a cut-off point between slow (<0.3) and rapid acetylators (>or=0.3). Twenty-seven samples belonged to the slow (mean 0.13; range: 0.03-0.25), 11 to the rapid (mean: 0.41; range: 0.34-0.48) acetylators. LightCycler analyses revealed 11 different NAT2 variant combinations, whereby *5B/*5B and *5B/*6A or *5A/*6C (each 21%), were the most frequent. The deduced acetylation status of the seven NAT2 SNPs matched perfectly with the 38 results determined by phenotyping. This study showed a 100% concordance between NAT2 phenotypes and the deduced NAT2 genotypes and the suitability of the high-speed NAT2-specific LightCycler analysis in a Caucasian population.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Acetylation
  • Adult
  • Arylamine N-Acetyltransferase / genetics*
  • Arylamine N-Acetyltransferase / metabolism
  • Biotransformation / genetics
  • Caffeine / metabolism*
  • Caffeine / pharmacokinetics
  • Genotype
  • Humans
  • Male
  • Middle Aged
  • Phenotype
  • Polymerase Chain Reaction
  • Polymorphism, Single Nucleotide*
  • Uracil / analogs & derivatives
  • Uracil / urine
  • Uric Acid / analogs & derivatives
  • Uric Acid / urine
  • White People
  • Xanthines / urine

Substances

  • Xanthines
  • Uric Acid
  • 1-methyluric acid
  • Caffeine
  • Uracil
  • 1-methylxanthine
  • 5-acetylamino-6-formylamino-3-methyluracil
  • Arylamine N-Acetyltransferase
  • NAT2 protein, human
  • 5-acetylamino-6-amino-3-methyluracil
  • 1,7-dimethyluric acid