Preimplantation genetic diagnosis for Down syndrome pregnancy

Preimplantation genetic diagnosis for Down syndrome pregnancy first-author corresp ‡Corresponding Author ‡Corresponding Author †E-mail:huanghefg@hotmail.com †E-mail:huanghefg@hotmail.com huanghefg@hotmail.com Department of Reproductive Endocrinology, Women’s Hospital, School of Medicine, Zhejiang University, Hangzhou 310006, China Department of Reproductive Endocrinology, Women’s Hospital, School of Medicine, Zhejiang University, Hangzhou 310006, China Objective: 2 To evaluate the effect of preimplantation genetic diagnosis (PGD) conducted for women who had Down syndrome pregnancy previously. Methods: 2 Trisomy 21 was diagnosed by using fluorescence in site hybridization (FISH) before embryo transfer in two women who had Down syndrome pregnancies. Each received one or two PGD cycles respectively. Results: 2 Case 1: one PGD cycle was conducted, two oocytes were fertilized and biopsied. One embryo is of trisomy 21 and the other of monosomy 21. No embryo was transferred. Case 2: two PGD cycles were conducted, in total, sixteen oocytes were fertilized and biopsied. Four embryos were tested to be normal, six of trisomy 21, and one of monosomy 21. Five had no signal. Four normal embryos were transferred but no pregnancy resulted. Conclusion: 2 For couples who had pregnancies with Down syndrome previously, PGD can be considered, and has been shown to be an effective strategy. Down syndrome Fluorescence in site hybridization (FISH) Preimplantation genetic diagnosis (PGD) INTRODUCTION 1 Down syndrome (DS), also named 21 trisomy syndrome, is a chromosome disorder associated either with an extra chromosome 21 or an effective trisomy for chromosome 21. The population risk for Down syndrome is approximately 1 in 800 live births (Roizen and Patterson, 2003 2003 ) and the recurrence risk after the birth of an affected child is approximately 1%~2% (Mikkelsen and Stene, 1979 1979 ; Daniel et al., 1982 1982 ). At present, women who had history of Down syndrome pregnancy may give birth to a normal baby with the help of prenatal diagnosis. However, they may suffer repeated pregnancy terminations following Down syndrome conception and these couples are seeking genetic diagnosis before implantation to avoid subsequent pregnancy termination (Conn et al., 1999 1999 ). Preimplantation genetic diagnosis (PGD) is a technique that was originally developed as an alternative to prenatal diagnosis for couples at high risk of transmitting a genetic defect. It involves the screening of in vitro fertilization (IVF) or intracytoplasm sperm injection (ICSI) generated embryos by the genetic analysis of one or two biopsied blastomeres, allows scientists to check specific genetic defects of the embryo so that only embryos not affected by the tested disease or balanced for the tested chromosomes will be transferred (Sermon et al., 2005 2005 ). Compared to the traditional methods of prenatal diagnosis, PGD offers genetic analysis at the earliest stage in fetal development, leading to the avoidance of abnormal pregnancy and subsequent pregnancy termination. Fluorescence in site hybridization (FISH) is the technique of choice for detecting the chromosome status in single cells for PGD, using fluorochrome-labelled DNA probes that are complementary to DNA sequences specific to individual chromosomes. It permits sexing the embryos (in case of X-linked diseases), simultaneous enumeration of up to nine chromosomes for aneuploidy screening (for the detection of abnormal numbers of chromosomes) and structural chromosome abnormalities (such as unbanlanced translocations) (Rubio et al., 2005a 2005a ). FISH has been proved to be a valuable tool for the study of aneuploidy in early human embryos (Schrurs et al., 1993 1993 ; Munné et al., 1998c 1998c ; 2003 2003 ). The number of cycles of PGD for aneuploidy screening (PGS) (Sermon et al., 2005 2005 ) has been steadily increasing. However, not many reports of PGD for specific single chromosome 21 were available. Here, we report two cases whose state of chromosome 21 was detected by using FISH in preimplantation embryos and who had Down syndrome pregnancy. At the same time, the present situation of preimplantation genetic diagnosis used for Down syndrome is discussed. SUBJECTS AND METHODS 1 Ethical approval 2 The clinical application of PGD was licensed by the Health Ministry of People’s Republic of China. The protocol was approved by local Ethical Committee and informed written consent was obtained from both patients. Patients 2 Case 1: The couple was referred for PGD after three abnormal pregnancies (one spontaneous abortion, a Down syndrome child and one termination of Down syndrome pregnancy). Maternal age was 43 years. Case 2: The couple was referred for PGD after two Down syndrome conceptions (one termination of pregnancy and a Down syndrome child). Maternal age was 30 years. Both couples had normal karyotype as proven by G-banded metaphase chromosome analysis of cultured peripheral blood lymphocytes. Both couples were counseled about the PGD procedure and the need to confirm diagnosis with prenatal diagnosis. Materials and methods 2 Ovarian stimulation, fertilization and embryo culture: ICSI was conducted as described by van Steirteghem et al.( 1993 1993 ) to avoid sperm contamination. Briefly, patients underwent superovulation and oocyte retrieval was conducted by vaginal ultrasound guided aspiration. Oocytes were subjected to ICSI and assessed for normal fertilization after 17 h. Embryos were cultured in vitro until 44 h post-insemination when the majority had reached the 6~10 cell stage. Biopsy and slide preparation: On day 3 the embryos biopsy procedure was performed after decompaction in Ca/Mg-free Scandinavian Embryo Medium (Science Scandinavia) and acidified Tyrode’s solution for zona drilling. One blastomere was biopsied from each embryo. After biopsy, embryos were immediately returned to normal culture conditions. Each single blastomere was transferred to poly-L-lysine slides in spreading solution (0.01 mol/L HCl, 0.1% Tween 20) that was gently agitated until lysis occurred and all nuclei were clear of cytoplasm. The slides were left to air dry for ~20 min, washed in PBS for 5 min and dehydrated through an ethanol series (70%, 80%, 100% at 5 min each). FISH: FISH was conducted as Munné et al.( 1998a 1998a ) described. Biopsied blastomeres were hybridized with probe of locus-specific indicator (LSI) 21 (SpectrumOrange) provide by Vysis Inc. (Illinois, USA) and LSI (13q34) (SpectrumGreen) was used as control. FISH signals were visualized using an Olympus-AX fluorescence microscope. Only embryos with normal chromosomal complement were transferred on day 4 of culture while those exhibiting chromosomal abnormalities were excluded. RESULTS 1 Case 1 2 One PGD cycle was conducted. Three oocytes were obtained, of which two were fertilized normally and biopsied on day 3. FISH results showed that one is of trisomy 21, and the other is of monosomy 21. No embryo was transferred (Figs. 1a and 1b 1a and 1b ). Case 2 2 Two PGD cycles were conducted. In the first cycle, ten oocytes were obtained, of which eight were fertilized normally and biopsied on day 3. Two gave a normal/balanced signal pattern, and were transferred on day 4. Three embryos were proved to be trisomy 21, and one of monosomy 21. The other two embryos were inconclusive because of hybridization failure. No pregnancy resulted. In the second cycle, seventeen oocytes were obtained, fourteen were fertilized normally, and eight biopsied on day 3. Two gave a normal/balanced signal pattern, and were transferred on day 4. Three embryos were proved to be trisomy 21. The other three embryos were inconclusive. No pregnancy resulted (Figs. 1c and 1d 1c and 1d ). DISCUSSION 1 Down syndrome is classically characterized by birth defects and mental retardation. In conventional prenatal diagnosis, definitive diagnosis of Down syndrome requires amniocentesis, chorionic villus sampling or umbilical blood sampling. Efforts have been made to reduce the risk of prenatal diagnosis, including obtaining fetal cells from maternal blood (Ho et al., 2003 2003 ; Yang et al., 2003 2003 ; 2006 2006 ), applying molecular genetic techniques to broaden the range of identifiable genetic disorders (Solassol et al., 2003 2003 ; Yang et al., 2006 2006 ) and raising the detection rate of noninvasive prenatal screening to reduce the false-positive rate in the hope of reducing the examine rate of invasive procedure (Cicero et al., 2003 2003 ; Muller et al., 2003 2003 ; Nicolaides, 2004 2004 ; Spencer et al., 2003 2003 ). However, invasive diagnosis is unavoidably combined with some complications and there are many religious controversies over whether to terminate the affected pregnancy. Moreover, the termination of pregnancy in the event of Down syndrome is not acceptable for some couples and there is growing demand for PGD diagnosis of Down syndrome. PGD is an alternative to prenatal diagnosis combined with assisted reproductive technology (ART) and genetic diagnosis technology, which offers genetic analysis at the earliest stage in fetal development, allows unaffected embryos to be identified and transferred to the uterus (Munné et al., 2004 2004 ). PGD avoids not only pregnancy termination when the fetus is proved affected by prenatal diagnosis, but also potential complications of invasive procedures, such as abortion, hemorrhage and uterus infection. Moreover, PGD can solve the political and ethical controversy of whether to terminate the affected pregnancy. Since Handyside et al.( 1990 1990 ) reported the first established pregnancy using PGD in 1990, the PGD cycles have been stably increasing worldwide (Geraedts et al., 2000 2000 ; Harper et al., 2006 2006 ; Sermon et al., 2005 2005 ), and PGD has been extensively applied in sex-linked disorders, single gene disease, autosomal abnormalities, and aneuploid detection of women with advanced age. Therefore, PGD has been a practical option for avoiding the birth of affected children, representing an important complement to traditional prenatal diagnosis (Kuliev and Verlinsky, 2004 2004 ). PGS was applied primarily to infertile women with the following indications: advanced maternal age (AMA) (Kahraman et al., 2000 2000 ), recurrent implantation failure (RIF) (Caglar et al., 2005 2005 ; Taranissi et al., 2005 2005 ) and recurrent miscarriage (RM) (Rubio et al., 2005b 2005b ). PGD for numerical chromosome abnormalities has four potential benefits: (1) to prevent trisomic offspring by analyzing chromosomes 13, 16, 18, 21 and 22 (Kuliev et al., 2003 2003 ); (2) to reduce spontaneous abortions (Munné et al., 1998b 1998b ; Rubio et al., 2003 2003 ); (3) to reduce multiple pregnancy by minimizing the number of embryos necessary for replacement and successful pregnancy (Munné, 2002 2002 ); and (4) to improve implantation. Many cycles of PGS were performed worldwide using FISH probes for chromosome 13, 18, 21 and others. However, only 4 reports of 5 cases of PGD for specific chromosome 21 are available (Conn et al., 1998 1998 ; 1999 1999 ; Scriven et al., 2001 2001 ; Luo et al., 2002 2002 ). All cases had history of Down syndrome pregnancy. PGD was conducted using cleavage stage embryo biopsy and FISH analysis for chromosome 21. Each couple underwent one or two treatment cycles. The case information and results are summarized in Table 1 1 . Probes: Locus-specific indicator (LSI) 13 and 21 Probes: LSI 21 and a biotinylated 14q subtelomere probe NP: No pregnancy; OHC: One healthy child; ABP: A biochemical pregnancy 2 Reference 2 Case 2 Maternal age 2 Pregnancy times (DS pregnancies) 2 Parental karyotypes 2 Cycle 3 Oocytes 2 Biopsy diagnosis 2 Transferred 2 Outcome Collected Fertilized Biopsied 8 Conn et al. ( 1998 1998 ) 8 1 a a 8 31 8 4 (3) 8 45, XX, der (13; 21) (q10; q10) 4 1 4 11 4 8 4 6 2 normal Yes 8 NP 2 monosomy 21, 13 No 1 monosomy 21 No 1 monosomy 13 No 4 2 4 14 4 10 4 5 2 monosomy 21, 13 No 1 tetrasomy 21 No 1 trisomy 21 No 1 unconclusive No 7 Conn et al. ( 1999 1999 ) 3 2 3 36 3 4 (3) 3 Normal 1 3 31 (total) 3 13 (total) 2 2 trisomy 21 No 3 NP 2 2 2 3 2 normal Yes 1 trisomy 21 No 4 3 4 32 4 5 (1) 4 46, XX, t (6;21) (q13; 22.3) 3 1 3 32 (total) 3 11 (total) 3 4 1 normal Yes 3 ABP 2 trisomy 21 No 1 monosomy 21 No 2 0 9 Scriven et al.( 2001 2001 ) 9 4 b b 9 39 9 2 (1) 9 45, XX, der (14; 21) (q10; q10) 3 1 3 11 3 7 3 5 3 normal Yes 3 NP 1 normal No 1 trisomy 21, 14 No 6 2 6 15 6 10 6 9 3 normal Yes 6 OHC 1 trisomy 21 No 1 monosomy 21 No 1 trisomy 14 No 1 monosomy 14 No 2 unconclusive No 4 Luo et al. ( 2002 2002 ) 4 5 4 30 4 1 (1) 4 Normal 4 1 4 14 4 13 4 8 3 normal Yes 4 OHC 3 normal No 1 trisomy 21 No 1 no signal No 9 Author’s cases 2 6 (case 1) 2 43 2 3 (2) 2 Normal 2 1 2 3 2 2 2 2 1 trisomy 21 No 1 monosomy 21 No 7 7 (case 2) 7 30 7 2 (2) 7 Normal 4 1 4 10 4 8 4 8 2 normal Yes 4 NP 3 trisomy 21 No 1 monosomy 21 No 2 unconclusive No 3 2 3 17 3 14 3 8 2 normal Yes 3 NP 3 trisomy 21 No 3 unconclusive No The PGD cycles summarized here were conducted for women who had pregnancies of Down syndrome previously. In total of above summarized reports and ours, 60 embryos were biopsied. Of them, only 22 (36.7%) were tested to be normal for chromosome 21, 18 (30.0%) transferred, 2 clinical pregnancies resulted, and two healthy children were born and no Down syndrome pregnancy occurred. In addition, the FISH results of surplus embryos (not biopsied because of fewer than six cells on day 3) showed that the abnormality rate was as high as 81.8% (18/22) (Conn et al., 1998 1998 ; 1999 1999 ; Cozzi et al., 1999 1999 ; Scriven et al., 2001 2001 ; Luo et al., 2002 2002 ). These data indicate that applying PGD for couples with history of Down syndrome effectively avoided affected pregnancy, although the pregnancy rate is low. The data here also imply that successful pregnancy has more possibility occurring in couples with high rate of normal embryos (50% and more), as suggested by Scriven et al.( 2001 2001 ), Conn et al.( 1998 1998 ; 1999 1999 ) and Luo et al.( 2002 2002 ). Women who have a history of Down syndrome pregnancy are at high risk of recurrent Down syndrome (Hook, 1992 1992 ). Data from livebirths and from amniocenteses in Europe (Stene et al., 1984 1984 ; Warburton et al., 1987 1987 ) showed that for women with a history of trisomy 21, there is an increased recurrence risk of trisomy 21, but not other trisomy. Although a wider selection of commercial probes is now becoming available, the development of suitable probe combinations for detecting multiple chromosome abnormality is expensive (Conn et al., 1999 1999 ). Therefore, for patients with a history of Down syndrome pregnancy, applying PGD for chromosome 21 is practicable (Scriven et al., 2001 2001 ; Luo et al., 2002 2002 ). The cause of recurrent Down syndrome is complex. Although studies have shown that most of recurrent trisomy 21 pregnancies may be the result of chance alone (Panaqalos et al., 1992 1992 ), the possibility of parental translocation and gonadal mosaicism has important implications for recurrence risk. Other evidence suggests that women with a diminished ovarian reserve may have a higher risk of a trisomic conception, so that biological ovarian age may be a better indicator of trisomy risk than chronological age (van Montfrans et al., 2001 2001 ). Recently, many suggestions for the mechanisms of non-age-related risks (absolute excess risk) have been put forward, indicating that women who experienced a Down syndrome pregnancy at a young age have a greater absolute excess risk of recurrence than those whose first Down syndrome pregnancy was at an older age (Morris et al., 2005 2005 ). Thus, the causative reasons for high recurrence of Down syndrome in our cases and in Luo et al.( 2002 2002 )’s in which both parents had a normal chromosome complement are needed for further analysis. In conclusion, PGD for trisomy 21 with FISH avoids affected pregnancy and offers favorable results. Although standard prenatal diagnosis remains the method of choice for aneuploidy, for couples who had pregnancies with Down syndrome previously, PGD can also be considered, and has been shown to be an effective strategy. References GS Caglar , B Asimakopoulos , N Nikolettos , K Diedrich , S Al-Hasani . Preimplantation genetic diagnosis for aneuploidy screening in repeated implantation failure . Reprod Biomed Online , 2005 . 10 ( 3 ): 381 - 388 . 15820047 . Preimplantation genetic diagnosis for aneuploidy screening in repeated implantation failure S Cicero , R Bindra , G Rembouskos , K Spencer , KH Nicolaides . Integrated ultrasound and biochemical screening for trisomy 21 using fetal nuchal translucency, absent fetal nasal bone, free beta-hCG and PAPP-A at 11 to 14 weeks . Prenat Diagn , 2003 . 23 ( 4 ): 306 - 310 . 12673635 DOI: 10.1002/pd.588 http://doi.org/10.1002/pd.588 . Integrated ultrasound and biochemical screening for trisomy 21 using fetal nuchal translucency, absent fetal nasal bone, free beta-hCG and PAPP-A at 11 to 14 weeks CM Conn , JC Harper , RM Winston , JD Delhanty . Infertile couples with Robertsonian translocations: preimplantation genetic analysis of embryos reveals chaotic cleavage divisions . Hum Genet , 1998 . 102 ( 1 ): 117 - 123 . 9490289 DOI: 10.1007/s004390050663 http://doi.org/10.1007/s004390050663 . Infertile couples with Robertsonian translocations: preimplantation genetic analysis of embryos reveals chaotic cleavage divisions CM Conn , J Cozzi , JC Harper , RM Winston , JD Delhanty . Preimplantation genetic diagnosis for couples at high risk of Down syndrome pregnancy owing to parental translocation or mosaicism . J Med Genet , 1999 . 36 ( 1 ): 45 - 50 . 9950365 . Preimplantation genetic diagnosis for couples at high risk of Down syndrome pregnancy owing to parental translocation or mosaicism J Cozzi , CM Conn , J Harper , RM Winston , M Rindl , PA Farndon , JD Delhanty . A trisomic germ cell line and precocious chromatid segregation leads to recurrent trisomy 21 conception . Hum Genet , 1999 . 104 ( 1 ): 23 - 28 . 10071188 DOI: 10.1007/s004390050905 http://doi.org/10.1007/s004390050905 . A trisomic germ cell line and precocious chromatid segregation leads to recurrent trisomy 21 conception A Daniel , L Stewart , T Saville , R Brookwell , H Paul , S Purvis-Smith , PR Lam-Po-Tang . Prenatal diagnosis in 3000 women for chromosome X-linked and metabolic disorders . Am J Med Genet , 1982 . 11 ( 1 ): 61 - 75 . 7065004 DOI: 10.1002/ajmg.1320110109 http://doi.org/10.1002/ajmg.1320110109 . Prenatal diagnosis in 3000 women for chromosome X-linked and metabolic disorders J Geraedts , A Handyside , J Harper , I Liebaers , K Sermon , C Staessen , A Thornhill , S Viville , L Wilton . ESHRE PGD Consortium Steering Committee: ESHRE Preimplantation Genetic Diagnosis (PGD) Consortium: data collection II (May 2000) . Hum Reprod , 2000 . 15 ( 12 ): 2673 - 2683 . 11098044 . ESHRE PGD Consortium Steering Committee: ESHRE Preimplantation Genetic Diagnosis (PGD) Consortium: data collection II (May 2000) AH Handyside , EH Kontogianni , K Hardy , RM Winston . Pregnancies from boipsied human preimplantation embryos sexed by Y-specific DNA amplification . Nature , 1990 . 344 ( 6268 ): 768 - 770 . 2330030 DOI: 10.1038/344768a0 http://doi.org/10.1038/344768a0 . Pregnancies from boipsied human preimplantation embryos sexed by Y-specific DNA amplification JC Harper , K Boelaert , J Geraedts , G Harton , WG Kearns , C Moutou , N Muntjewerff , S Repping , S SenGupta , PN Scriven . ESHRE PGD consortium data collection V: cycles form January to December 2002 with pregnancy follow-up to October 2003 . Hum Reprod , 2006 . 21 ( 1 ): 3 - 21 . 16172150 DOI: 10.1093/humrep/dei292 http://doi.org/10.1093/humrep/dei292 . ESHRE PGD consortium data collection V: cycles form January to December 2002 with pregnancy follow-up to October 2003 SS Ho , K O′Donoghue , M Choolani . Fetal cells in maternal blood: state of the art for non-invasive prenatal diagnosis . Annu Acad Med Singapore , 2003 . 32 ( 5 ): 597 - 603 . quiz 604 . Fetal cells in maternal blood: state of the art for non-invasive prenatal diagnosis EB Hook . DJH Brock , CH Rodeck , MA Ferguson-Smith . Prevalence, Risks and Recurrence . Prenatal Diagnosis and Screening , 1992 . : Edinburgh Churchill Livingston , 351 - 392 . . Prevalence, Risks and Recurrence S Kahraman , M Bahce , H Samli , N Imirzalioglu , K Yakisn , G Cengiz , E Donmez . Healthy births and ongoing pregnancies obtained by preimplantation genetic diagnosis in patients with advanced maternal age and recurrent implantation failure . Hum Reprod , 2000 . 15 ( 9 ): 2003 - 2007 . 10967004 DOI: 10.1093/humrep/15.9.2003 http://doi.org/10.1093/humrep/15.9.2003 . Healthy births and ongoing pregnancies obtained by preimplantation genetic diagnosis in patients with advanced maternal age and recurrent implantation failure A Kuliev , Y Verlinsky . Thirteen years’ experience of preimplantation diagnosis: report of the Fifth International Symposium on Preimplantation Genetics . Reprod Biomed Online , 2004 . 8 ( 2 ): 229 - 235 . 14989805 . Thirteen years’ experience of preimplantation diagnosis: report of the Fifth International Symposium on Preimplantation Genetics A Kuliev , J Cieslak , Y Ilkevitch , Y Verlinsky . Chromosomal abnormalities in a series of 6733 human oocytes in preimplantation diagnosis for age-related aneuploidies . Reprod Biomed Online , 2003 . 6 ( 1 ): 54 - 59 . 12626143 . Chromosomal abnormalities in a series of 6733 human oocytes in preimplantation diagnosis for age-related aneuploidies C Luo , FQ Xing , SQ Luo . Normal pregnancy in a woman having had a child with trisomy 21 syndrome before: application of preimplantation genetic diagnosis . Di Yi Jun Yi Da Xue Xue Bao , 2002 . 22 ( 3 ): 269 - 271 . (in Chinese) 12390787 . Normal pregnancy in a woman having had a child with trisomy 21 syndrome before: application of preimplantation genetic diagnosis M Mikkelsen , J Stene . JD Murken , S Stengel-Rutkwski , E Schwinger . Previous Child with Down Syndrome and Other Chromosome Aberrations . Proceedings of the 3rd European Conference on Prenatal Diagnosis of Genetic Disorders , 1979 . : Dtuggart Enke , 22 - 29 . . Previous Child with Down Syndrome and Other Chromosome Aberrations JK Morris , DE Mutton , E Alberman . Recurrences of free trisomy 21: analysis of data from the National Down Syndrome Cytogenetic Register . Prenat Diagn , 2005 . 25 ( 12 ): 1120 - 1128 . 16231400 DOI: 10.1002/pd.1292 http://doi.org/10.1002/pd.1292 . Recurrences of free trisomy 21: analysis of data from the National Down Syndrome Cytogenetic Register F Muller , C Benattar , F Audibert , N Roussel , S Dreux , H Cuckle . First-trimester screening for Down syndrome in France combining fetal nuchal translucency measurement and biochemical markers . Prenat Diagn , 2003 . 23 ( 10 ): 833 - 836 . 14558029 DOI: 10.1002/pd.700 http://doi.org/10.1002/pd.700 . First-trimester screening for Down syndrome in France combining fetal nuchal translucency measurement and biochemical markers S Munné . Preimplantation genetic diagnosis of numerical and structural chromosome abnormalities . Reprod Biomed Online , 2002 . 4 ( 2 ): 183 - 196 . 12470583 . Preimplantation genetic diagnosis of numerical and structural chromosome abnormalities S Munné , J Fung , MJ Cassel , C Marquez , HU Weier . Preimplantation genetic analysis of translocations: case-specific probes for interphase cell analysis . Hum Genet , 1998 . 102 ( 6 ): 663 - 674 . 9703428 DOI: 10.1007/s004390050759 http://doi.org/10.1007/s004390050759 . Preimplantation genetic analysis of translocations: case-specific probes for interphase cell analysis S Munné , C Magli , M Bahce , J Fung , M Legator , L Morrison , J Cohert , L Gianaroli . Preimplantation diagnosis of the aneuploidies most commonly found in spontaneous abortions and live births: XY, 13, 14, 15, 16, 18, 21, 22 . Prenat Diagn , 1998 . 18 ( 13 ): 1459 - 1466 . 9949446 DOI: 10.1002/(SICI)1097-0223(199812)18:13<1459::AID-PD514>3.0.CO;2-V http://doi.org/10.1002/(SICI)1097-0223(199812)18:13<1459::AID-PD514>3.0.CO;2-V . Preimplantation diagnosis of the aneuploidies most commonly found in spontaneous abortions and live births: XY, 13, 14, 15, 16, 18, 21, 22 S Munné , C Marquez , A Reing , J Garrisi , M Alikani . Chromosome abnormalities in embryos obtained after conventional in vitro fertilization and intracytoplasmic sperm injection . Fertil Steril , 1998 . 69 ( 5 ): 904 - 908 . 9591501 DOI: 10.1016/S0015-0282(98)00039-9 http://doi.org/10.1016/S0015-0282(98)00039-9 . Chromosome abnormalities in embryos obtained after conventional in vitro fertilization and intracytoplasmic sperm injection S Munné , M Sandalinas , T Escudero , E Velilla , R Walmsley , S Sadowy , J Cohen , D Sable . Improved implantation after preimplantation genetic diagnosis of aneuploidy . Reprod Biomed Online , 2003 . 7 ( 1 ): 91 - 97 . 12930584 . Improved implantation after preimplantation genetic diagnosis of aneuploidy S Munné , T Escudero , P Colls , Z Xuezhong , M Oter , M Garrisi , F Barnes , C Zouves , L Werlin , C Maqli , 等 . Predictability of preimplantation genetic diagnosis of aneuploidy and translocations on prospective attempts . Reprod Biomed Online , 2004 . 9 ( 6 ): 645 - 651 . 15670413 . Predictability of preimplantation genetic diagnosis of aneuploidy and translocations on prospective attempts KH Nicolaides . Nuchal translucency and other first-trimester sonographic markers of chromosomal abnormalities . Am J Obstet Gynecol , 2004 . 191 ( 1 ): 45 - 67 . 15295343 DOI: 10.1016/j.ajog.2004.03.090 http://doi.org/10.1016/j.ajog.2004.03.090 . Nuchal translucency and other first-trimester sonographic markers of chromosomal abnormalities CG Panaqalos , CCJr Talbot , JG Lewis , PA Adelsberqer , MB Petersen , JL Serre , MO Rethore , MC de Blois , P Parent , AA Schinzel . DNA polymorphism analysis in families with recurrence of free trisomy 21 . Am J Hum Genet , 1992 . 51 ( 5 ): 1015 - 1027 . 1415248 . DNA polymorphism analysis in families with recurrence of free trisomy 21 NJ Roizen , D Patterson . Down’s syndrome . Lancet , 2003 . 361 ( 9365 ): 1281 - 1289 . 12699967 DOI: 10.1016/S0140-6736(03)12987-X http://doi.org/10.1016/S0140-6736(03)12987-X . Down’s syndrome C Rubio , C Simon , F Vidal , L Rodrigo , J Pehlivan , J Remohi , A Pellicer . Chromosomal abnormalities and embryo development in recurrent miscarriage couples . Hum Reprod , 2003 . 18 ( 1 ): 182 - 188 . 12525464 DOI: 10.1093/humrep/deg015 http://doi.org/10.1093/humrep/deg015 . Chromosomal abnormalities and embryo development in recurrent miscarriage couples C Rubio , L Rodrigo , I Perez-Cano , A Mercader , E Mateu , P Buendia , J Remohi , C Simon , A Pellicer . FISH screening of aneuploidies in preimplantation embryos to improve IVF outcome . Reprod Biomed Online , 2005 . 11 ( 4 ): 497 - 506 . 16274616 . FISH screening of aneuploidies in preimplantation embryos to improve IVF outcome C Rubio , T Pehlivan , L Rodrigo , C Simon , J Remohi , A Pellicer . Embryo aneuploidy screening for unexplained recurrent miscarriage: a minireview . Am J Reprod Immunol , 2005 . 53 ( 4 ): 159 - 165 . 15760376 DOI: 10.1111/j.1600-0897.2005.00260.x http://doi.org/10.1111/j.1600-0897.2005.00260.x . Embryo aneuploidy screening for unexplained recurrent miscarriage: a minireview BM Schrurs , RM Winston , AH Handyside . Preimplantation diagnosis of aneuploidy using fluorescent in situ hybridization: evaluation using a chromosome 18-specifid probe . Hum Reprod , 1993 . 8 ( 2 ): 296 - 301 . 8473437 . Preimplantation diagnosis of aneuploidy using fluorescent in situ hybridization: evaluation using a chromosome 18-specifid probe PN Scriven , FA Flinter , PR Braude , CM Ogilvie . Robertsonian translocations reproductive risks and indications for preimplantation genetic diagnosis . Hum Reprod , 2001 . 16 ( 11 ): 2267 - 2273 . 11679502 DOI: 10.1093/humrep/16.11.2267 http://doi.org/10.1093/humrep/16.11.2267 . Robertsonian translocations reproductive risks and indications for preimplantation genetic diagnosis K Sermon , C Moutou , J Harper , J Geraedts , P Scriven , L Wilton , MC Magli , A Michiels , S Viville , C de Die . ESHRE PGD consortium data collection IV: May-December 2001 . Hum Reprod , 2005 . 20 ( 1 ): 19 - 34 . 15550497 DOI: 10.1093/humrep/deh552 http://doi.org/10.1093/humrep/deh552 . ESHRE PGD consortium data collection IV: May-December 2001 J Solassol , H Rahil , V Sapin , D Lemery , B Dastugue , O Boespflug-Tanguy , I Creveaux . Detection of trisomy 21 by quantitative fluorescent-polymerase chain reaction in uncultured amniocytes . Prenat Diagn , 2003 . 23 ( 4 ): 287 - 291 . 12673631 DOI: 10.1002/pd.579 http://doi.org/10.1002/pd.579 . Detection of trisomy 21 by quantitative fluorescent-polymerase chain reaction in uncultured amniocytes K Spencer , CE Spencer , M Power , C Dawson , KH Nicolaides . Screening for chromosomal abnormalities in the first trimester using ultrasound and maternal serum biochemistry in a one-stop clinic: a review of three years prospective experience . BJOG , 2003 . 110 ( 3 ): 281 - 286 . 12628268 DOI: 10.1046/j.1471-0528.2003.02246.x http://doi.org/10.1046/j.1471-0528.2003.02246.x . Screening for chromosomal abnormalities in the first trimester using ultrasound and maternal serum biochemistry in a one-stop clinic: a review of three years prospective experience J Stene , E Stene , M Mikkelsen . Risk for chromosome abnormality at amniocentesis following a child with a non-inherited chromosome aberration. A European Collaborative Study on Prenatal Diagnoses 1981 . Prenat Diagn , 1984 . 4 ( 7 ): 81 - 95 . 6235486 DOI: 10.1002/pd.1970040707 http://doi.org/10.1002/pd.1970040707 . Risk for chromosome abnormality at amniocentesis following a child with a non-inherited chromosome aberration. A European Collaborative Study on Prenatal Diagnoses 1981 M Taranissi , T El-Toukhy , A Gorgy , Y Verlinsky . Influence of maternal age on the outcome of PGD for aneuploidy screening in patients with recurrent implantation failure . Reprod Biomed Online , 2005 . 10 ( 5 ): 628 - 632 . 15949221 . Influence of maternal age on the outcome of PGD for aneuploidy screening in patients with recurrent implantation failure JM van Montfrans , CB Lambalk , MHA van Hooff , JMG Vugt . Are elevated FSH concentrations in the pre-conceptional period a risk factor for Down’s syndrome pregnancies? . Hum Reprod , 2001 . 16 ( 6 ): 1270 - 1273 . 11387304 DOI: 10.1093/humrep/16.6.1270 http://doi.org/10.1093/humrep/16.6.1270 . Are elevated FSH concentrations in the pre-conceptional period a risk factor for Down’s syndrome pregnancies? AC van Steirteghem , Z Nagy , H Joris , J Liu , C Staessen , J Smitz , A Wisanto , P Devroey . High fertilization and implantation rates after intracytoplasmic sperm injection . Hum Reprod , 1993 . 8 ( 7 ): 1061 - 1066 . 8408487 . High fertilization and implantation rates after intracytoplasmic sperm injection D Warburton , J Kline , Z Stein , M Hutzler , A Chin , T Hassold . Does the karyotype of a spontaneous abortion predict the karyotype of a subsequent abortion? Evidence from 273 women with two karyotyped sponstaneous abortions . Am J Hum Genet , 1987 . 41 ( 3 ): 465 - 483 . 3631080 . Does the karyotype of a spontaneous abortion predict the karyotype of a subsequent abortion? Evidence from 273 women with two karyotyped sponstaneous abortions YH Yang , SH Kim , ES Yang , SK Kim , IK Kim , YW Park , JS Cho , YH Lee . Prenatal diagnosis of fetal trisomy 21 from maternal peripheral blood . Yonsei Med J , 2003 . 44 ( 2 ): 181 - 186 . 12728455 . Prenatal diagnosis of fetal trisomy 21 from maternal peripheral blood YH Yang , ES Yang , JY Kwon , IK Kim , YW Park . Prenatal diagnosis of trisomy 21 with fetal cells in maternal blood using comparative genomic hybridization . Fetal Diagn Ther , 2006 . 21 ( 1 ): 125 - 133 . 16354990 DOI: 10.1159/000089062 http://doi.org/10.1159/000089062 . Prenatal diagnosis of trisomy 21 with fetal cells in maternal blood using comparative genomic hybridization 10.1631/jzus.2007.B0515 Project supported by the National Basic Research Program of China (Nos. 2006CB944006 and 2006CB504004), the Key Research Program of Zhejiang Province, China (No. 2006C13078) and the Bureau of Science and Technology of Hangzhou, China (No. 20061123B03) 2006-08-25 2007-02-13 2022-01-05 Copyright © 2007, Journal of Zhejiang University Science 2007 Journal of Zhejiang University-SCIENCE B (Biomedicine and Biotechnology) 07 08