标记21和13号染色体双色FISH探针及其在快速产前诊断中的应用  

Labeling of the 21 and 13 Chromosomal Enumeration Dual Color FISH Probes and Their Efficiency in Rapid Prenatal Screen of the Aneuploidy

作　　者：滕奔琦[1] 章钧[1] 范建辉[1] 尹玉竹[1] 王静 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]达安基因诊断中心,广东广州510080

出　　处：《中山大学学报（医学科学版）》2010年第6期884-888,共5页Journal of Sun Yat-Sen University:Medical Sciences

基　　金：广东省医学科研基金(B2009076);广东省科技计划项目(2009B060700107)

摘　　要：【目的】采用细菌人工染色体(BAC)克隆分别建立21和13号染色体的直标型荧光原位杂交(FISH)探针,并分析其在产前快速诊断21/13号染色体数目异常的可行性。【方法】通过生物信息学网站挑选位于21q22.2(DSCR)和13q14(RB基因)的BAC克隆,培养并提取DNA后采用切口平移法进行红/绿色(21/13)荧光的直接标记。取上述探针与中期细胞进行杂交,鉴定探针的杂交位置。取10例羊水标本分别采用标记的FISH探针(实验组)与商品化探针(对照组)进行检测,分别计算探针的杂交率与准确率,采用随机化配对设计资料的t检验分析两种探针杂交效能的差异。取2008年10月至2010年4月期间,来我院行羊膜腔穿刺术的453例羊水标本作为实验组进行上述探针的FISH分析,同时对该453例标本亦进行细胞培养核型分析作为对照,并将两组检测结果进行比较,验证探针灵敏度、特异度及阴、阳性预测价值。【结果】采用BAC克隆标记的FISH探针荧光信号清晰、明确,与核型分析对照位置吻合;杂交率及准确率与商品化探针相比不存在统计学差异;对453例羊水标本21/13三体进行检测的灵敏度为83.3%(5/6),特异度为100%(447/447),阳性预测价值为100%(5/5),阴性预测价值为99.8%(447/448)。【结论】采用BAC克隆标记建立的21/13号染色体直标型FISH探针荧光信号清晰、明确,结果准确可靠、成本低廉,可以用于未培养羊水标本的产前快速诊断21/13三体综合征。【Objective】 To investigate the methods of labeling 21 / 13 enumeration FISH probes from bacterial artificial chromosome （BAC） clones and their efficiency in rapid prenatal aneuploidy screen. 【Methods】 Two BAC clones from 21q22.2 （DSCR） and 13q14 were picked out from online bioinformatics databases and labeled of red / green to act as enumeration FISH probes of the chromosome 21 and 13. The hybridization positions of the probes were validated on metaphase cells. Ten amniotic fluid samples were simultaneously hybridized with commercial probes and home made probes to compare the hybridization rate and accuracy with t test. Probes efficiency was analyzed with 453 amniotic fluid samples gathered from November 2008 to April 2010 against the karyotyping results. 【Results】 FISH probes labeled from BAC clones hybridized to the exact place with excellent signal intensity,accuracy,sensitivity （83.3% ,5 / 6） specificity （100% ,447 / 447）,positive predictive value （100% ,5 / 5） and negative predictive value （99.8%,447 / 448）. 【Conclusion】 BAC clone FISH probes are cost-effective tools in rapid prenatal screen of 21 / 13 aneuploidy with high insensitivity and specificity.

关 键 词：21三体综合征 13三体综合征 荧光原位杂交 

分 类 号：R714.53[医药卫生—妇产科学]