[The application of denaturing high performance liquid chromatography in detection of novel polymorphisms in
Xu-dong Wang1, Yan Fu, Hui-jie Jiang
1Emergency Department, Affiliated Beijing Tongren Hospital, Capital University of Medical Science, Beijing, 100730 PR China. wangxd5812@hotmail.com
Objective:
To detect single nucleotide polymorphisms(SNPs) in codon region of plasminogen activator inhibitor-1 (PAI-1) gene and associate them with development of coronary artery disease(CAD).
Methods:
Denaturing high performance liquid chromatography(DHPLC) and DNA sequencing were used to scan all of exons in PAI-1 gene in 93 CAD patients and 123 control subjects.
Results:
Two polymorphisms in exon 2, G43A and G49A, which results in missense mutation Ala15Thr and Val17Ile, were found. Heterozygotes for these variants as well as wild type homozygotes were only found in all subjects. The association of genotypes with CAD development and with PAI 1 antigen level were evaluated. The comparison of genotype frequency distribution between two groups revealed that there were more heterozygous carriers of mutation at both polymorphic sites in control than in CAD subjects, but there was no statistically significant difference between them. And genotypes of both polymorphisms were not associated with PAI-1 level.
Conclusion:
Two SNPs in exon 2 of PAI-1 gene were identified by DHPLC method. Both polymorphisms were not associated with the incidence of CAD.
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