21三体综合症植入前诊断的临床前运用研究  

Preclinical Study of Preimplantation Genetic Diagnosis of 21 Trisomy Syndrome

作　　者：徐伟[1] 陈华丽[1] 曹友德[1] 袁友红[1] 李灼日[1] 

机构地区：[1]湖南省人民医院,中国湖南长沙410005

出　　处：《生命科学研究》2006年第4期342-345,共4页Life Science Research

基　　金：湖南省科技计划项目(科技攻关04sk3031)

摘　　要：探讨建立一个高效、稳定的21三体遗传病植入前诊断的方法,以染色体G显带核型分析为对照,取正常成人外周血单个淋巴细胞40枚、21三体患者外周血单个淋巴细胞40枚及单卵裂球20枚,采用荧光定量PCR技术同时扩增21号染色体上特异区域基因片段(DSCR)和12号染色体上管家基因(GAPDH)片断作内对照,结果在正常组织同时扩增二片断的有效率为95%(38/40),扩增产物的荧光强度比值为1.00±0.05;21三体患者单个淋巴细胞同时扩增二片断的有效扩增率为92.5%(37/40),DSCR/GAPDH荧光强度的比值约为1.58±0.17;单卵裂球的扩增效率为80.0%(16/20).三者实验结果与染色体核型分析结果完全一致,准确率100%.研究结果表明荧光定量PCR技术产前检测21三体综合征具有准确、快速、安全、实用等特点,有较高的临床推广应用价值.To develop a rapid, high efficient, reliable method for preimplantation genetic diagnosis of 21 trisomy syndrome, fluorescence quantitative PCR method was used to detect the original copies of Down syndrome by amplifying the critical region gene （DSCR3） and house-keeping gene GAPDH. Then the ratio of DSCR3/ GAPDH was calculated , which can directly detect the additional copy of chromosome 21 by comparing simultaneously amplified two genes. The samples were from 40 clinically diagnosed Down syndrome patients, 40 normal controls and 20 donated embryos. The amplification efficiencies of single lymphocyte fluorescence PCR of trisomy 21 Syndrome group, control group and blastomere group were 92. 5 %, 95 % and 80% respectively. The ratios of DSCR3/ GAPDH products were 1.58 ±0. 17 and 1.00 ±0. 05 （ x^- ± S） for trisomy 21 and controls respectively. The difference between these two groups was statistically significant （ P〈 0.01 ）. The results of fluorescence PCR products were the same as those of the chromosome analysis. Fluorescence quantitative PCR analysis revealed that fluorescence quantitative polymerase chain reaction is a practical, safe, precise and rapid preimplantation diagnosis approach to detect 21 trisomy syndrome.

关 键 词：荧光定量PCR 单细胞 植入前诊断 

分 类 号：R394[医药卫生—医学遗传学]