聚合酶链式反应-变性高效液相色谱技术在唐氏综合征筛查中的应用

Application of PCR-DHPLC technology in Down's syndrome screening

目的:探讨聚合酶链式反应-变性高效液相色谱技术(polymerase chain reaction-denaturing high performance liquidchromatography,PCR-DHPLC)在唐氏综合征筛查中的应用。方法:针对21号染色体上的基因座D21S1409、D21S1422、D21S1435和GATA163G03设计引物,PCR扩增唐氏综合征患儿样本和正常样本,DHPLC仪检测;同步进行外周血染色体核型分析,并以核型分析为金标准,评价PCR-DHPLC技术的准确性。结果:柱温50℃时,唐氏综合征患儿样本DHPLC检测结果呈现特异唐氏综合征峰。结论:PCR-DHPLC方法具有较高的敏感性和特异性,可用于唐氏综合征的高通量快速筛查。

OBJECTIVE：To evaluate the use of polymerase chain reaction-denaturing high performance liquid chromatography technology（PCR-DHPLC） in Down syndrome screening.METHODS：Primers of D21S1409, D21S1422,D21S1435 and GATA163G03 loci on chromosome 21 were designed.The samples of the children with Down＇s syndrome or the normal ones were amplified using PCR method.The PCR amplification products were identified by denaturing high performance liquid chromatography technology.At the same time the same samples were analyzed with karyotyping,to evaluate the accuracy of PCR-DHPLC.RESULTS：The column temperature was 50℃,the Down syndrome samples showed specific DHPLC peak of Down syndrome.CONCLUSION：The PCR-DHPLC method edmonstrated high sensitivity and specificity.It could be used in high-throughput rapid screening for Down＇s syndrome.