无创染色体筛查技术在体外受精-胚胎移植中的初步研究和临床应用
摘要
目的探讨应用无创染色体筛查(NICS)技术检测囊胚培养液中的游离染色体和应用胚胎活检技术检测胚胎染色体的一致性,以初步确定NICS的临床应用价值。方法收集2016年3-12月在该院行卵胞浆内单精子注射—胚胎移植(ICSI-ET)的患者15例,分为A组(子宫及子宫附件无异常)、B组(多囊卵巢综合征)、C组(子宫内膜异位症);获得的23枚囊胚分为新鲜胚胎组和卵裂期解冻胚胎组; NICS筛查作为观察组,胚胎植入前遗传学筛查(PGS)作为对照组,比较二者的一致性。通过对14例有不良妊娠史的不孕不育患者行NICS胚胎筛选,初步确定NICS的临床应用价值。结果观察组与对照组的吻合率为82. 61%,一致性检验Kappa值为0. 65,两者一致性一般。新鲜胚胎组与卵裂期解冻胚胎组的吻合率分别为84. 62%和80. 00%,两组Kappa值分别为0. 69和0. 58,提示两组的一致性均一般。A、B、C 3组的吻合率分别为87. 50%、80. 00%和80. 00%,3组的Kappa值分别为0. 83、0. 58和0. 54,提示A组一致性较好,B、C组一致性一般。14例有不良妊娠史的不孕不育患者中有7例正常妊娠,孕19周进行胎儿羊水染色体检测,确认NICS检测结果准确。结论不同病症的不孕不育患者、不同来源的胚胎NICS和PGS检测结果无差别。NICS和PGS具有同样的准确性和可靠性。但NICS可能具有比PGS更广泛的应用前景。
引文
[1]Battaglia DE,Goodwin P,Klein NA,et al.Influence of maternal age on meiotic spindle assembly in oocytes from naturally cycling women[J].Hum Reprod,1996,11(10):2217-2222.
[2]Scott RT,Jr,Upham KM,Forman EJ,et al.Blastocyst biopsy with comprehensive chromosome screening and fresh embryo transfer significantly increases in vitro fertilization implantation and delivery rates:A randomized controlled trial[J].Fertil Steril,2013,100(3):697-703.
[3]Keltz MD.Preimplantation genetic screening(PGS)with comparative genomic hybridization(CGH)following day3 single cell blastomere biopsy markedly improves IVF outcomes while lowering multiple pregnancies and miscarriages[J].J Assist Reprod Genet,2013,30(10):1333-1339.
[4]Cimadomo D,Capalbo A,Ubaldi FM,et al.The impact of biopsy on human embryo developmental potential during preimplantation genetic diagnosis[J].Biomed Res Int,2016,2016:7193075.
[5]Xu J,Fang R,Chen L,et al.Noninvasive chromosome screening of human embryos by genome sequencing of embryo culture medium for in vitro fertilization[J].Pnas,2016,113(42):11907-11912.
[6]Zong C,Lu S,Chapman AR,Xie XS.Genome-wide detection of single nucleotide and copy number variations of a single human cell[J].Science,2012,338(6114):1622-1626.
[7]Wu Y,Lv Z,Yang Y,et al.Blastomere biopsy influences epigenetic reprogramming during early embryo development,which impacts neural development and function in resulting mice[J].Cell Mol Life Sci,2014,71(9):1761-1774.
[8]Zhao HC,Zhao Y,Li M,et al.Aberrant epigenetic modification in murine brain tissues of offspring from preimplantation genetic diagnosis blastomere biopsies[J].Biol Reprod,2013,89(5):117-124.
[9]Palini S,Galluzzi L,De Stefani S,et al.Genomic DNA in human blastocoeie fluid[J].Reprod Biomed Online,2013,26(6):603-610.
[10]Gianaroli L,Magli MC,Pomante A,et al.Blastocentesis:a source of DNA for preimplantation genetic testing results from a pilot study[J].Fertil Steri,2014,102(6):1692-1699.
[11]Stigliani S,Anserini P,Venturini PL,et al.Mitochondrial DNAcontent in embryo culture medium is significantly associated with human embryo fragmentation[J].Hum Reprod,2013,28(10):2652-2660.
[12]Stigliani S,Persico L,Lagazio C,et al.Mitochondrial DNA in day3 embryo culture medium is a novel,non-invasive biomarker of blastocyst potential and implantation outcome[J].Mol Hum Reprod,2014,20(12):1238-1246.
[13]Wu H,Ding C,Shen X,et al.Medium-based noninvasive preimplantation genetic diagnosis for humanα-thalassemias-SEA[J].Medicine(Baltimore),2015,94(12):669-776.
[14]Huang L,Ma F,Chapman A,et al.Single-cell whole-genome amplification and sequencing:methodology and applications[J].Annu Rev Genomics Hum Genet,2015,16:79-102.
[15]Taylor TH,Gitlin SA,Patrick JL,et al.The origin,mechanisms,incidence and clinical consequences of chromosomal mosaicism in humans[J].Hum Reprod Update,2014,20(4):571-581.
[2]Scott RT,Jr,Upham KM,Forman EJ,et al.Blastocyst biopsy with comprehensive chromosome screening and fresh embryo transfer significantly increases in vitro fertilization implantation and delivery rates:A randomized controlled trial[J].Fertil Steril,2013,100(3):697-703.
[3]Keltz MD.Preimplantation genetic screening(PGS)with comparative genomic hybridization(CGH)following day3 single cell blastomere biopsy markedly improves IVF outcomes while lowering multiple pregnancies and miscarriages[J].J Assist Reprod Genet,2013,30(10):1333-1339.
[4]Cimadomo D,Capalbo A,Ubaldi FM,et al.The impact of biopsy on human embryo developmental potential during preimplantation genetic diagnosis[J].Biomed Res Int,2016,2016:7193075.
[5]Xu J,Fang R,Chen L,et al.Noninvasive chromosome screening of human embryos by genome sequencing of embryo culture medium for in vitro fertilization[J].Pnas,2016,113(42):11907-11912.
[6]Zong C,Lu S,Chapman AR,Xie XS.Genome-wide detection of single nucleotide and copy number variations of a single human cell[J].Science,2012,338(6114):1622-1626.
[7]Wu Y,Lv Z,Yang Y,et al.Blastomere biopsy influences epigenetic reprogramming during early embryo development,which impacts neural development and function in resulting mice[J].Cell Mol Life Sci,2014,71(9):1761-1774.
[8]Zhao HC,Zhao Y,Li M,et al.Aberrant epigenetic modification in murine brain tissues of offspring from preimplantation genetic diagnosis blastomere biopsies[J].Biol Reprod,2013,89(5):117-124.
[9]Palini S,Galluzzi L,De Stefani S,et al.Genomic DNA in human blastocoeie fluid[J].Reprod Biomed Online,2013,26(6):603-610.
[10]Gianaroli L,Magli MC,Pomante A,et al.Blastocentesis:a source of DNA for preimplantation genetic testing results from a pilot study[J].Fertil Steri,2014,102(6):1692-1699.
[11]Stigliani S,Anserini P,Venturini PL,et al.Mitochondrial DNAcontent in embryo culture medium is significantly associated with human embryo fragmentation[J].Hum Reprod,2013,28(10):2652-2660.
[12]Stigliani S,Persico L,Lagazio C,et al.Mitochondrial DNA in day3 embryo culture medium is a novel,non-invasive biomarker of blastocyst potential and implantation outcome[J].Mol Hum Reprod,2014,20(12):1238-1246.
[13]Wu H,Ding C,Shen X,et al.Medium-based noninvasive preimplantation genetic diagnosis for humanα-thalassemias-SEA[J].Medicine(Baltimore),2015,94(12):669-776.
[14]Huang L,Ma F,Chapman A,et al.Single-cell whole-genome amplification and sequencing:methodology and applications[J].Annu Rev Genomics Hum Genet,2015,16:79-102.
[15]Taylor TH,Gitlin SA,Patrick JL,et al.The origin,mechanisms,incidence and clinical consequences of chromosomal mosaicism in humans[J].Hum Reprod Update,2014,20(4):571-581.