建立非重复序列FISH探针及在21/13三体产前诊断中的应用  

Labeling and evaluation of non-repeat FISH probes of 21/13 enumeration in prenatal diagnosis

作　　者：章钧[1] 李佩琼[2] 尹玉竹[1] 滕奔琦[1] 叶燕绸[1] 侯红瑛[1] 

机构地区：[1]中山大学附属第三医院产科,广东广州510630 [2]广州医学院基础学院医学遗传与细胞生物学教研室,广东广州510182

出　　处：《分子诊断与治疗杂志》2011年第6期368-372,共5页Journal of Molecular Diagnostics and Therapy

基　　金：广东省科技计划项目(2009B060700107);广东省科技计划项目(2010B060900031);广东省医学科学基金(B2009076)

摘　　要：目的建立21/13号染色体的非重复序列荧光原位杂交(FISH)探针,比较其与商品化含重复序列的探针在对羊水脱落细胞进行快速产前诊断21/13三体的效能方面的差异。方法分析DSCR区域和RB基因区域DNA序列,剔除重复序列后进行PCR扩增和荧光标记,制备21/13号染色体FISH计数探针,与G显带中期淋巴细胞杂交,鉴定探针的杂交位置。选取50例孕周为18～35周的孕妇,经羊膜腔穿刺取羊水标本,分别在培养前后采用Cytocell的21/13号染色体FISH计数探针和自制的非重复序列FISH计数探针进行杂交,计算探针杂交率,并以核型分析结果作为参考,计算探针的准确率。两组数据间差异采用t检验进行统计学分析。结果非重复序列探针位置准确,荧光信号明确,背景更低;两种探针与培养后细胞杂交的杂交率和准确率无明显差异;但对于未培养的羊水间期细胞,非重复序列探针杂交的准确率明显优于普通探针,杂交率两者相似。结论采用非重复序列PCR方式制备21/13染色体计数探针用于羊水间期细胞杂交能明显改善探针的杂交效率和信噪比,有利于提高检测的准确度。Objective To label 21/13 enumeration FISH probes from non-repeat sequences through PCR amplification and to compare their efficacy with that of commercial FISH probes in prenatal diagnosis of trisomy 21/13. Methods The non-repeat portion of DSCR and RB gene sequences were amplified and labeled as enumeration FISH probes of chromosomes 21 and 13. The probes were then tested with G-banding karotyping slides. Fifty amniotic fluid samples with gestational weeks from 18-35 were parallelly tested with non-repeat probes and Cytocell 21/13 FISH probes. Hybridization rate and accuracy were compared between the two groups with t tests. Results Non-repeat probes exhibited excellent efficiency and low signal/background ratio in detecting arnniotic fluid samples, thus resulted in better accuracy. Conclusion FISH probes labeling from non-repeat sequences were more fit for prenatal diagnosis of trisomy 21/13 from amniotic fluid samples with better accuracy.

关 键 词：荧光原位杂交 产前诊断 

分 类 号：R714.5[医药卫生—妇产科学]