Mutation Screening of the AT-III Gene in Pregnant and Post-Partum Women With Venous Thromboembolism by DHPLC

Background: AT-III gene mutations are associated with venous thromboembolism (VTE) in pregnant women or puerperants. In this study we established a denaturing highperformance liquid chromatography (DHPLC) assay to screen mutation/polymorphism of the AT-III gene. Methods: Forty pregnant and post-partum women with venous VTE and 50 normal pregnant and post-partum women were enrolled. Ten pairs of primers were designed on the AT-III gene, which flank the promoter region and exons 1-6, including their exon/ intron boundaries. Serial DHPLC analyses were performed under the optimal conditions for each amplicon. Results: Two mutations and 6 single nucleotide polymorphisms (SNPs) were detected in the case group, including a novel silent mutation in exon 4 and a reported pathogenic mutation in exon 6 (G13328A). Two of the 6 SNPs detected in this screen are novel alterations. Conclusion: Denaturing high-performance liquid chromatography assay is a highly sensitive, time saving, and less labor-intensive method for the mutation/polymorphism detection of the AT-III gene.