open access
Detection of mutation in NAT II gene as a method of determination of izoniazyd (INH) acetylation type in human population
open access
Abstract
Introduction: Isoniazid (INH) is a drug widely used in the treatment of tuberculosis. INH is metabolized to acetylisoniazid by N-acetyltransferase (NAT) in the liver. The rate of INH acetylation is genetically determined. NAT isozymes are encoded at 2 loci; one encodes NAT1, formerly known as the monomorphic form of the enzyme, while the other encodes the polymorphic NAT2, which is responsible for individual differences in the ability to acetylate certain compounds. The objective of the present study was to apply the genotyping of the fast and slow acetylators for personalized therapeutic dose.
Material and methods: Plasma concentrations of INH were determined with biological method in the authors modification. This method warrants high accuracy and secured repeatable results. Genomic DNA was isolated from the blood samples. DNA extracted by Blood DNA Kit and amplified by PCR by Spurr [1] with two primers. PCR product was cut separately with 4 different restriction enzymes: Dde1, Kpn1, Tag1, and BamH1.
Results: Four different NAT 2 alleles were detected in the study population. The presence of any 2 mutant alleles defines the slow-acetylator genotype, whereas rapid acetylators have 1 or 2 wild-type NAT2*4 alleles.
Conclusion: On the basis of our results we suggest the using of NAT 2 genotyping for discrimination of the fast and slow acetylators in monitoring of tuberculosis therapy.
Abstract
Introduction: Isoniazid (INH) is a drug widely used in the treatment of tuberculosis. INH is metabolized to acetylisoniazid by N-acetyltransferase (NAT) in the liver. The rate of INH acetylation is genetically determined. NAT isozymes are encoded at 2 loci; one encodes NAT1, formerly known as the monomorphic form of the enzyme, while the other encodes the polymorphic NAT2, which is responsible for individual differences in the ability to acetylate certain compounds. The objective of the present study was to apply the genotyping of the fast and slow acetylators for personalized therapeutic dose.
Material and methods: Plasma concentrations of INH were determined with biological method in the authors modification. This method warrants high accuracy and secured repeatable results. Genomic DNA was isolated from the blood samples. DNA extracted by Blood DNA Kit and amplified by PCR by Spurr [1] with two primers. PCR product was cut separately with 4 different restriction enzymes: Dde1, Kpn1, Tag1, and BamH1.
Results: Four different NAT 2 alleles were detected in the study population. The presence of any 2 mutant alleles defines the slow-acetylator genotype, whereas rapid acetylators have 1 or 2 wild-type NAT2*4 alleles.
Conclusion: On the basis of our results we suggest the using of NAT 2 genotyping for discrimination of the fast and slow acetylators in monitoring of tuberculosis therapy.
Keywords
N-acetyltransferase; isoniazid; polymorphism NAT II; treatment of tuberculosis
About this articleTitle
Detection of mutation in NAT II gene as a method of determination of izoniazyd (INH) acetylation type in human population
Journal
Advances in Respiratory Medicine
Issue
Pages
134-139
Published online
2007-06-03
Bibliographic record
Pneumonol Alergol Pol 2007;75(2):134-139.
Keywords
N-acetyltransferase
isoniazid
polymorphism NAT II
treatment of tuberculosis
Authors
Ewa Augustynowicz-Kopeć
Anna Zabost
Monika Kozińska
Sylwia Brzezińska
Zofia Zwolska
