微阵列芯片比较基因组杂交技术在植入前遗传学诊断中的应用研究  

A preliminary study on the application of array comparative genomic hybridization for preimplantation genetic diagnosis

作　　者：谢言信[1] 徐艳文[1] 苗本郁[1] 曾艳红[1] 周灿权[1] 

机构地区：[1]中山大学附属第一医院生殖医学中心,广州510080

出　　处：《中华医学遗传学杂志》2013年第3期283-287,共5页Chinese Journal of Medical Genetics

基　　金：基金项目：广东省科技基金（2008A030201028）

摘　　要：目的建立适用于植入前遗传学诊断的微阵列芯片比较基因组杂交技术（arraycomparativegenomichybridization，arrayCGH）。方法对以下3种来源的细胞进行arrayCGH分析：（1）经胰酶消化的核型分别为46，XX、46，XY、47，XX，＋13的B2、C38、A1的3株人胚胎干细胞，分离后获取3～5个细胞；（2）在本中心进行体外受精与胚胎移植（in-vitrofertilization—embryotransfer，IVF—ET）的正常受精来源的废弃胚胎卵裂球；（3）源自5对罗氏或相互易位携带者的夫妻、在本中心进行植入前遗传学诊断的10个废弃胚胎，其中8个胚胎经荧光原位杂交（fluorescenceinsituhybridization，FIsH）诊断为异常，1枚无信号，l枚诊断正常但胚胎发育停滞。经全基因组扩增后，对扩增产物进行24sureV3或24sure＋芯片检测，采用BlueFuseMulti软件进行数据分析。结果（1）对来自B2、C38、A1系干细胞株的3～5个细胞进行arrayCGH分析，结果与核型鉴定一致；（2）对来自2枚正常受精胚胎的6个卵裂球的扩增产物行24sureV3分析，1枚胚胎的2个卵裂球分别为非整倍体和正常核型，另1枚胚胎的4个卵裂球中2个正常，另外2个均为-22，＋13。用24sure＋重复上述实验，其结果与前一致；（3）对来自染色体易位行植入前遗传学诊断的10枚胚胎进行arrayCGH分析，4枚胚胎在arrayCGH分析时与FISH诊断结果相符，其中2枚FISH与arrayCGH结果完全一致，2枚通过arrayCGH分析发现除FISH发现的异常外的其他多条染色体数目异常。1枚胚胎FISH检测无信号，而arrayCGH诊断为13三体。5枚与FISH诊断结果不符的胚胎中，1枚来自罗氏易位FISH诊断为13单体，两种arrayCGH芯片均诊断为14三体及其他多条染色体非整倍体改变，1枚胚胎碎片多且发育停滞（该易位患者另2枚胚胎诊断均相符），其余3枚结果不相符的胚胎均来源于染色体（p13；q11）的相互易位。结论应用arrayCGHObjective To assess the value of array comparative genomic hybridization （array CGH） technique for preimplantation genetic diagnosis （PGD）. Methods Array CGH was performed on three types of cells, which included 3-5 cells isolated from B2/C38/A1 embryonic stem cell lines, single cells isolated from two discarded normal fertilized embryos, and 10 blastocysts biopsied from 5 couples undergoing PGD for chromosomal translocations. For the 10 blastocysts, 8 were abnormal embryos, 1 appeared to be normal but showed arrested development, and 1 embryo was without any fluorescence signals. 24sure V3 or 24sure ＋ array chips were applied for CGH analysis. The results were analyzed with a BlueFuse Multi software. Results （1） The results of cells from B2/C3/A1 embryo stem cells by array CGH were consistent with karyotyping analysis. （2） For the 6 single cell samples from two discarded embryos, 2 blastomeres from one embryo were diagnosed as with aneuploidy and a normal karyotype, respectively. Two out of 4 blastomeres biopsied from another embryo were normal, whilst the remaining two were diagnosed with aneuploidies of --22 and ＋13. Repeated detection with 24sure＋ array was consistent with the 24sure V3 result. （3） Ten cell masses from 10 embryos in PGD cycles were successfully analyzed with array CGH, among which four were confirmed with fluorescence in situ hybridization （FISH） on day 3. In two of them, array CGH confirmed FISH diagnosis. For the remaining two, additional aneuploidies for chromosomes not tested byFISH were discovered by array CGH. Another embryo diagnosed as no signal by FISH was found to have trisomy 13 by array CGH. The remaining 5 embryos also showed discordant results by FISH and array CGH. One embryo from a Robertsonian translocation carrier was found to have monosomy 1a by FISH but trisomy 14 and additional aneuploidies by both 24sure V3 and 24sure q- chips. One embryo with many fragments and arrested development by D5 showed discordant results by FISH and array CGH. Howe

关 键 词：植入前遗传学诊断 微阵列芯片比较基因组杂交技术 荧光原位杂交技术 罗氏易位 相互易位 非整倍体 

分 类 号：R394[医药卫生—医学遗传学]