利用双色实时荧光定量PCR法快速进行产前诊断

Rapid diagnosis of trisomy 21 or trisomy 18 by dual real-time fluorescence relative quantitative PCR

目的利用双色实时荧光定量PCR法进行21-三体与18-三体的快速产前诊断。方法分别以21号的APP基因与18号染色体的TYMS基因为目的基因设计引物和不同荧光标记的Taqman探针,以相对定量指标△CT值判断21号与18号染色体的数目;并将其检测结果与传统染色体核型分析方法进行对比。结果 60例羊水细胞区分出了4例21-三体,3例18三体标本,正常人的△CT值为-0.89±0.22,21-三体患者及18-三体患者的△CT值分别-0.04±0.17和-1.55±0.24,患者与正常人标本组之间无交叉重叠,均有明显差异(P<0.001)。结论实验建立的双重实时荧光定量PCR能用于快速诊断羊水细胞的21-三体和18-三体等非整倍体。

Objective： To establish a dual real - time fluorescence relative quantitative PCR method for diagnosis of trisomy 21 and trisomy 18. Methods： The APP gene on chromosome 21 and TYMS gene on chromosome 18 were used as the target genes. The primers and probes were design for the two genes, and which were amplified in a single reaction. The △CT value of relative quantitative was used to differentiate trisomy 21 and trisomy 18 patients from normal individual. Results ： the PCR product ratio of APP to TYMS, in the normal control the ACT was -0. 89 ±0. 22; in trisomy 21 and trisomy 18 were -0. 04 ±0. 17 and - 1.55 ±0. 24, respectively. The △CT value from trisomy 21 or trisomy 18 group were dramatically different from normal group （P 〈0. 001 ）. Conclusion： Real - time fluorescence quantitative PCR is a rapid and accurate method for prenatal diagnosis of trisomy 21 and trisomy 18.