title: Single step PCR for detection of allelic variation of MDR1 gene (P-glycoprotein) among three ethnic groups in Malaysia.
abstract: BACKGROUND
  P-glycoprotein (PgP) is the most extensively studied ATP-binding cassette (ABC) coded by MDR1 gene. To date, 29 single nucleotide polymorphisms (SNPs) have been identified; but only SNP C3435T has been correlated with intestinal PgP expression levels and shown to influence the absorption of orally taken drugs that are PgP substrates. Individuals homozygous for the T allele have more than fourfold lower PgP expression compared with C/C individuals. We developed a one step primer based allele specific PCR method to detect SNP at C3435T to investigate the distribution of this genotype in the local population.
  
  
  METHODS
  DNA was extracted from 5 mL of whole blood using standard salting-out method. Primers were designed specific to 3' end which amplify the variants of C3435T. The method was validated by direct DNA sequencing. Seven hundred and sixty-three healthy blood donors comprising of three major ethnic groups in Malaysia were recruited and DNA subjected to genotyping of C3435T using this method.
  
  
  RESULTS
  The method was found to be robust and reproducible in detecting SNP of C3435T. Interethnic variations in genotype and allele frequency were observed in PgP among the ethnic groups. In comparison to both the Caucasians and the other Asian countries, the Malay and Chinese showed a higher frequency of allele C (50-60%); while the Indian exhibits a lower frequency (40%), similar to other Indian populations.
  
  
  CONCLUSIONS
  Using a new simple method to investigate the distribution of C3435T, we found that the allele frequency of MDR1 showed variablity between the different ethnic groups within the Malaysian population.
authors.affiliation: Faculty of Pharmacy, Universiti Teknologi MARA, 41050 Shah Alam, Malaysia. tehlaykek@salam.uitm.edu.my
authors.forename: L K
authors.initials: LK
authors.lastname: Teh
authors.forename: W L
authors.initials: WL
authors.lastname: Lee
authors.forename: J
authors.initials: J
authors.lastname: Amir
authors.forename: M Z
authors.initials: MZ
authors.lastname: Salleh
authors.forename: R
authors.initials: R
authors.lastname: Ismail
mesh_terms: D020168:ATP Binding Cassette Transporter, Subfamily B, Member 1 / Q000235:genetics*; D000483:Alleles; D044466:Asian People / Q000208:ethnology / Q000235:genetics; D001483:Base Sequence; D004247:DNA / Q000737:chemistry / Q000235:genetics; D005260:Female; D005787:Gene Frequency; D014644:Genetic Variation; D005838:Genotype; D006801:Humans; D008296:Malaysia; D008297:Male; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000379:methods*; D011110:Polymorphism, Genetic; D015203:Reproducibility of Results; D017422:Sequence Analysis, DNA; D044465:White People / Q000208:ethnology / Q000235:genetics
chemical_list: D020168:ATP Binding Cassette Transporter, Subfamily B, Member 1; D004247:DNA