应用同源基因定量PCR方法快速检测Down综合征

Rapid detection of Down's syndrome by homologous gene quantitative PCR

本文设计一对引物 ,用聚合酶链反应同时扩增位于 2 1号染色体的人肝型磷酸果糖激酶基因 (PFKL -CH2 1)以及与其高度同源的位于 1号染色体上人肌型磷酸果糖激酶基因 (PFKM -CH1) ,然后同光密度扫描进行定量比较分析 ,结果显示 ,在 31例正常人这两个同源基因的扩增产量比值 0 .939± 0 .16 1;而 16例Down综合征患者的比值约为 1.5 89± 0 .16 4,两组相比有高度显著性差异 (P <0 .0 0 1) ,正常组与异常组的比值分布不存在重叠区 ,阴阳性结果易于判断 ,因此这种同源基因定量聚合酶联反应有可能发展成一种新的快速诊断Down综合征的方法。

Objective:To study rapid gene diagnosis of trisomy 21.Methods:Use a method termed the homologous gene quantitative polymerase chain reaction (HGQ-PCR),which designs one pair of primers and which can directly identify the additional copy of chromosome 21 by simultaneiously amplfying two highl homologous genes of the human liver-type phosphofructokinase located on chromosome 21 (PFKL-CH21) and the human musle-type phosphofructokinase located on chromosome (PFKM-CH1)for self detecting determination.Results:On analysis of 31 normal individuals and 16 cases of Down's syndrome,the relative ratio of PFKL-CH21/PFKM-CH1 product was 0.993±0.161(mean±SD)for disomy DNA and 1.589±0.164 for trisomy DNA,respectively.The difference between these two groups was highly significan(P<0.001)Conclusion:These results shows that this HGQ-PCR method is reliable and may be used for a rapid prenatal gene diagnosis of Down's syndrome.