中华医学遗传学杂志
中華醫學遺傳學雜誌
중화의학유전학잡지
CHINESE JOURNAL OF MEDICAL GENETICS
2013年
3期
283-287
,共5页
谢言信%徐艳文%苗本郁%曾艳红%周灿权
謝言信%徐豔文%苗本鬱%曾豔紅%週燦權
사언신%서염문%묘본욱%증염홍%주찬권
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
植入前遺傳學診斷%微陣列芯片比較基因組雜交技術%熒光原位雜交技術%囉氏易位%相互易位%非整倍體
식입전유전학진단%미진렬심편비교기인조잡교기술%형광원위잡교기술%라씨역위%상호역위%비정배체
Preimplantation genetic diagnosis%Array comparative genomic hybridization%Fluorescence in situ hybridization%Robertsonian translocation%Reciprocal translocation%Aneuploidy
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(array comparative genomic hybridization,array CGH).方法 对以下3种来源的细胞进行array CGH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-vitro fertilization-embryo transfer,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(fluorescence in situ hybridization,FISH)诊断为异常,1枚无信号,1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24sure V3或24sure+芯片检测,采用BlueFuse Multi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array CGH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24sure V3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array CGH分析,4枚胚胎在array CGH分析时与FISH诊断结果相符,其中2枚FISH与array CGH结果完全一致,2枚通过array CGH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array CGH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array CGH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array CGH可同时完成对胚胎染色体结构检测及全染色体组非整倍体筛查,但在断裂点距离着丝粒位置较近的染色体1区的易位应选择分辨率高的24sure+芯片.
目的 建立適用于植入前遺傳學診斷的微陣列芯片比較基因組雜交技術(array comparative genomic hybridization,array CGH).方法 對以下3種來源的細胞進行array CGH分析:(1)經胰酶消化的覈型分彆為46,XX、46,XY、47,XX,+13的B2、C38、A1的3株人胚胎榦細胞,分離後穫取3～5箇細胞；(2)在本中心進行體外受精與胚胎移植(in-vitro fertilization-embryo transfer,IVF-ET)的正常受精來源的廢棄胚胎卵裂毬；(3)源自5對囉氏或相互易位攜帶者的伕妻、在本中心進行植入前遺傳學診斷的10箇廢棄胚胎,其中8箇胚胎經熒光原位雜交(fluorescence in situ hybridization,FISH)診斷為異常,1枚無信號,1枚診斷正常但胚胎髮育停滯.經全基因組擴增後,對擴增產物進行24sure V3或24sure+芯片檢測,採用BlueFuse Multi軟件進行數據分析.結果 (1)對來自B2、C38、A1繫榦細胞株的3～5箇細胞進行array CGH分析,結果與覈型鑒定一緻；(2)對來自2枚正常受精胚胎的6箇卵裂毬的擴增產物行24sure V3分析,1枚胚胎的2箇卵裂毬分彆為非整倍體和正常覈型,另1枚胚胎的4箇卵裂毬中2箇正常,另外2箇均為-22,+13.用24sure+重複上述實驗,其結果與前一緻；(3)對來自染色體易位行植入前遺傳學診斷的10枚胚胎進行array CGH分析,4枚胚胎在array CGH分析時與FISH診斷結果相符,其中2枚FISH與array CGH結果完全一緻,2枚通過array CGH分析髮現除FISH髮現的異常外的其他多條染色體數目異常.1枚胚胎FISH檢測無信號,而array CGH診斷為13三體.5枚與FISH診斷結果不符的胚胎中,1枚來自囉氏易位FISH診斷為13單體,兩種array CGH芯片均診斷為14三體及其他多條染色體非整倍體改變,1枚胚胎碎片多且髮育停滯(該易位患者另2枚胚胎診斷均相符),其餘3枚結果不相符的胚胎均來源于染色體(p13;q11)的相互易位.結論 應用array CGH可同時完成對胚胎染色體結構檢測及全染色體組非整倍體篩查,但在斷裂點距離著絲粒位置較近的染色體1區的易位應選擇分辨率高的24sure+芯片.
목적 건립괄용우식입전유전학진단적미진렬심편비교기인조잡교기술(array comparative genomic hybridization,array CGH).방법 대이하3충래원적세포진행array CGH분석:(1)경이매소화적핵형분별위46,XX、46,XY、47,XX,+13적B2、C38、A1적3주인배태간세포,분리후획취3～5개세포；(2)재본중심진행체외수정여배태이식(in-vitro fertilization-embryo transfer,IVF-ET)적정상수정래원적폐기배태란렬구；(3)원자5대라씨혹상호역위휴대자적부처、재본중심진행식입전유전학진단적10개폐기배태,기중8개배태경형광원위잡교(fluorescence in situ hybridization,FISH)진단위이상,1매무신호,1매진단정상단배태발육정체.경전기인조확증후,대확증산물진행24sure V3혹24sure+심편검측,채용BlueFuse Multi연건진행수거분석.결과 (1)대래자B2、C38、A1계간세포주적3～5개세포진행array CGH분석,결과여핵형감정일치；(2)대래자2매정상수정배태적6개란렬구적확증산물행24sure V3분석,1매배태적2개란렬구분별위비정배체화정상핵형,령1매배태적4개란렬구중2개정상,령외2개균위-22,+13.용24sure+중복상술실험,기결과여전일치；(3)대래자염색체역위행식입전유전학진단적10매배태진행array CGH분석,4매배태재array CGH분석시여FISH진단결과상부,기중2매FISH여array CGH결과완전일치,2매통과array CGH분석발현제FISH발현적이상외적기타다조염색체수목이상.1매배태FISH검측무신호,이array CGH진단위13삼체.5매여FISH진단결과불부적배태중,1매래자라씨역위FISH진단위13단체,량충array CGH심편균진단위14삼체급기타다조염색체비정배체개변,1매배태쇄편다차발육정체(해역위환자령2매배태진단균상부),기여3매결과불상부적배태균래원우염색체(p13;q11)적상호역위.결론 응용array CGH가동시완성대배태염색체결구검측급전염색체조비정배체사사,단재단렬점거리착사립위치교근적염색체1구적역위응선택분변솔고적24sure+심편.
Objective 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 ceils 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 by FISH 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 13 by FISH but trisomy 14 and additional aneuploidies by both 24sure V3 and 24sure + chips.One embryo with many fragments and arrested development by D5 showed discordant results by FISH and array CGH.However,the FISH and array CGH results for other two embryos from this reciprocal translocation carrier were consistent.Three embryos with inconsistent results by FISH and array CGH had a chromosomal translocation involving q11 region.Conclusion Array CGH is useful for PGD applications for its capability to detect structural chromosomal abnormalities through screening of aneuploidies.However,the 24sure V3 array may not suit detection of translocations with breakpoints close to the q11 region of chromosomes.
