Allele-specific polymerase chain reaction for genotyping human cytochrome P450 2E1

Authors
Citation
T. Sohda, Allele-specific polymerase chain reaction for genotyping human cytochrome P450 2E1, J CL LAB AN, 13(5), 1999, pp. 205-208
Citations number
14
Categorie Soggetti
Medical Research Diagnosis & Treatment
Journal title
JOURNAL OF CLINICAL LABORATORY ANALYSIS
ISSN journal
08878013 → ACNP
Volume
13
Issue
5
Year of publication
1999
Pages
205 - 208
Database
ISI
SICI code
0887-8013(1999)13:5<205:APCRFG>2.0.ZU;2-T
Abstract
Allele-specific polymerase chain reaction (AS-PCR) was applied to investiga te the cytochrome P450 2E1 (CYP2E1) genotype. AS-PCR is a competitive multi plex PCR method in which PCR amplification is successfully performed only b y using the sequence of 3' oligonucleotide ends as a DNA template in order to obtain an absolutely complementary product. I was able to produce allele -specific primers whose 3' ends had the base specific to Pst I polymorphism located within the 5'-flanking region of the CYP2E1 gene. Electrophoresis of the products showed that bands derived from common PCR products, allele C1 and C2, were clearly separate from each other due to the difference in t he size of the products. I tested 102 unrelated Japanese individuals, and t he results of both restriction fragment length polymorphism (RFLP) by Pst I or Rsa I and direct sequencing were in complete agreement with those of AS -PCR. These results lead me to conclude that AS-PCR is a simple and useful technique for investigating CYP2E1 genotype. J. Clin. Lab. Anal. 13. 205-20 8, 1999. (C) 1999 Wiley-Liss, Inc.