- Email: [email protected]
Comparison of day-3 and day-5 array-CGH diagnosis for 24 chromosome aneuploidy screening in terms of accuracy
DESIGN: Retrospective. MATERIALS AND METHODS: A retrospective review was conducted of all embryos that underwent PGS by 23 chromosome SNP microarrays from 1/1/2010 to 1/25/2011. Patients underwent standard in vitro fertilization (IVF) and preimplantation genetics screening (PGS) due to primarily a history of > 2 spontaneous miscarriages. Embryo biopsy was performed at either the cleavage or blastocyst stage. Sample DNA was amplified and analyzed using HumanCytoSNP-12 DNA beadchips and GenomeStudio and KaryoStudio software. Parental DNA was analyzed and compared to DNA from embryonic cells to determine the presence of benign copy number variations. Embryos derived from parents with known translocations or inversions were excluded from the study. Binomial confidence intervals for proportions were calculated. RESULTS: 1,902 embryos from 236 clinical IVF cycles were tested. 37.2% (707/1902) of all embryos were euploid without DUP or DEL. Morphologic fragmentation on some embryos led to 9.8% (186/1902) embryos with no molecular diagnosis obtained. Of the remaining 1009 abnormal embryos, 5.7% (58/1009) had DUP or DEL with no aneuploidy and 27.4% (277/ 1009) had DUP or DEL coupled with aneuploidy. 66.8% (674/1009) had aneuploidy without DUP or DEL. In total, there were 470 DUP, 175 involving the p arm and 295 involving the q arm. In contrast, there were only 95 DEL, 35 involving the p arm and 60 involving the q arm. DUP and q arm abnormalities were significantly more commonly observed compared to DEL or p arm abnormalities (P< .05). CONCLUSION: DUP and q arm errors seem to be much more common than DEL and p arm errors in the developing embryo. Therefore, duplications or structural aberrations in the q arm might be more compatible with early embryo development than deletions or errors in the p arm.
O-73 Monday, October 17, 2011 05:00 PM HALVING THE MULTIPLE PREGNANCY RATE IN A PREIMPLANTATION GENETIC DIAGNOSIS PROGRAM BY ADOPTING A NEW BLASTOCYST FREEZING POLICY. L. Pepas, V. Bolton, Y. Khalaf, T. El-Toukhy. Asssisted Conception Unit, Guy’s and St Thomas’ Hospitals Trust, London, United Kingdom. OBJECTIVE: The implications of multiple pregnancy in patients with inherited genetic disorders are significant. Elective single embryo transfer (eSET) has been recommended for preimplantation genetic diagnosis (PGD) for inherited genetic disorders to reduce multiple pregnancy rates. As the safety and success of cryo-thawed PGD cycles has been demonstrated, we sought to evaluate effect of a stricter SET policy on the outcomes of our PGD programme. DESIGN: Retrospective study. MATERIALS AND METHODS: Since January 2006, patients who had two or more blastocysts suitable for transfer after biopsy and genetic analysis on Day 3 were offered eSET and cryopreservation of surplus expanded blastocyst(s). To increase the uptake of eSET further, the policy was changed in June 2009 to include cryopreservation of early blastocysts too. We compared the outcomes of 424 fresh PGD cycles performed between January 2006 and May 2009 prior to this change; to 279 cycles performed between June 2009 and December 2010. RESULTS: Embryo transfer was achieved in 71% and 73% of cycles in each group respectively (P¼0.68). There was no difference in mean age (33.6 3.9 and 32.8 4.1) and FSH values (6.8 2.1 and 6.7 2.1) between the two groups. Our elective SET rate increased from 44% to 61% (P¼0.0006) and blastocyst freezing increased from 28% to 43% (P¼0.0001). The mean number of embryos transferred decreased from 1.35 0.6 to 0.92 0.7 (P¼0.0001). Per cycle reaching embryo transfer, the pregnancy rate increased from 47% to 57% (P¼0.0230), the clinical pregnancy rate increased from 39% to 49% (P¼0.0274) and the ongoing pregnancy/live birth rate increased from 34% to 43% (P¼0.0305). The multiple pregnancy rate per clinical pregnancy halved from 13% (n ¼ 15) to 6% (n ¼ 7) (P¼0.117). CONCLUSION: Through adoption of a stricter elective SET policy, encouraged by the relaxation of criteria for blastocyst freezing, we halved our multiple pregnancy rate without compromising PGD treatment outcome. Supported by: This project was funded internally.
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Abstracts
O-74 Monday, October 17, 2011 05:15 PM FISH REANALYSIS OF INNER CELL MASS AND TROPHECTODERM SAMPLES OF PREVIOUSLY ARRAY-CGH SCREENED BLASTOCYSTS REVEALS HIGH ACCURACY OF DIAGNOSIS AND NO SIGN OF MOSAICISM OR PREFERENTIAL ALLOCATION. A. Capalbo, G. Wright, L. Themaat, T. Elliott, L. Rienzi, Z. P. Nagy. GENERA, Reproductive Medicine, Rome, Italy; Reproductive Biology Associates, Atlanta, GA. OBJECTIVE: To study the blastocyst genetic constitution (possible chromosomal mosaicism between ICM and TE and within TE), after isolating ICM by a novel method.and to evaluate array-CGH accuracy against FISH testing DESIGN: Cross-sectional study. MATERIALS AND METHODS: 32 blastocysts diagnosed as genetically abnormal by array-CGH were donated to research. ICM was isolated from the trophectoderm (TE) using a novel mechanical separation which was confirmed by KRT18 immunostaining. 24 of the 32 embryo had single or double aneuploidy and 8 aneusomic for unbalanced product of translocation or for long single arm deletions/duplications but with normal ploidy (by aCGH). After biopsy, individual samples were fixed by Carnoy method and analyzed with conventional 9-chromosomes FISH and locus specific probes. RESULTS: We reanalyzed 32 ICM and 96 TE samples (3 separate groups of cells from each TE of the same embryo). Looking at the single chromosome level, 28 out of 28 aneuploidies diagnosed by a-CGH were confirmed by FISH reanalysis both in ICM and TE samples resulting in a analytical sensitivity of 100%(95%CI 90.2-100). For almost all embryos, the reported aneuploidies were constitutive. As regard to chromosomes diagnosed as disomic, only 2 out of 248 (specificity 99.1 95%CI 97.4-99.8) were misdiagnosed by a-CGH, a trisomy 13 in an unbalanced embryo from reciprocal translocation carrier observed in a constitutive state and a mosaic nullisomy/monosomy 16 in a double aneuploid embryo. Consequently, only one out of 32 embryos was misdiagnosed as false negative. Only in one case we observed a confined trisomy 17 in all nuclei of a TE sample from a trisomic embryo for chromosome 13. In all other embryos ICM and TE samples were concordant and confirmed the original a-CGH diagnosis. CONCLUSION: Our data show a very high analytical accuracy of a-CGH to diagnose embryo chromosomal constitution. Concordance between ICM and TE was reported for all embryos analyzed, showing no sign of mosaicism or preferential allocation at blastocyst stage.
O-75 Monday, October 17, 2011 05:30 PM COMPARISON OF DAY-3 AND DAY-5 ARRAY-CGH DIAGNOSIS FOR 24 CHROMOSOME ANEUPLOIDY SCREENING IN TERMS OF ACCURACY. P. Mir, L. Rodrigo, E. Mateu, A. Cervero, J. Martın, C. Rubio. PGD Unit, IVI-Valencia, Valencia, Comunitat Valenciana, Spain. OBJECTIVE: To describe the clinical outcome on day-3 single cell arrayCGH (aCGH) and to compare day-3 and day-5 aCGH diagnosis DESIGN: Prospective study. MATERIALS AND METHODS: Day-3 aCGH diagnosis was performed on 24 patients. Indications for Preimplantation Genetic Screening (PGS) were: recurrent miscarriage, repetitive implantation failure, severe male factor and advanced maternal age. Mean female age was 38.8 2.9 years. A single cell from 122 day-3 embryos was amplified (SureplexÒ, BlueGnome, Cambridge, UK). DNA amplification, labelling, hybridisation, washing, scanning and data analysis (24sureÒplatform, BlueGnome) was performed in a 24-hour protocol. On day-5, 39 chromosomally abnormal embryos were reanalysed by FISH using probes for the altered chromosomes (Vysis Inc., Downers Grove, IL,USA). Moreover in 14 abnormal but good developing blastocysts, aCGH was performed on throphoectoderm biopsy and the remaining cells of the embryo were analysed by FISH. RESULTS: A successful amplification was obtained in 117 blastomeres (95.9%), being 74 embryos diagnosed as abnormal (63.2%); 30 of them (40.5%) had aneuploidies only for the chromosomes analysed in our FISH program (chromosomes 13, 15, 16, 17, 18, 21, 22, X, Y); 29 (39.2%) had aneuploidies also for other chromosomes; and 15 (20.3%) had aneuploidies only for other chromosomes. In 20 cycles euploid embryos were transferred (83.3%), a clinical pregnancy was observed in 15 patients (62.5% per cycle; 75% per transference) with an implantation rate of 54.3%.
Vol. 96., No.3, Supplement, September 2011
FISH re-analysis on day-5 confirmed day-3 aCGH diagnosis in 38 embryos (97.4%). aCGH on trophectoderm biopsy showed 93.3% (14/15) of concordance with day-3 aCGH. The discordant was chaotic aneuploid by aCGH on day-3; euploid by aCGH on trophectoderm biopsy but chaotic aneuploid by FISH re-analysis. CONCLUSION: PGS using aCGH is suitable on single blastomeres with successful amplification efficiency and reliable results. In this study, day-5 trophectoderm biopsy does not improve the accuracy of the diagnosis compared to day-3.
O-76 Monday, October 17, 2011 05:45 PM SIGNIFICANT DECREASE IN MISCARRIAGES AFTER PREIMPLANTATION GENETIC DIAGNOSIS (PGD) FOR RECURRENT PREGNANCY LOSS USING ARRAY COMPARATIVE GENOME HYBRIDIZATION (Array CGH). J. Grifo, S. Ghadir, B. Kaplan, C. A. Laskin, M. Glassner, S. Munne. NYU Fertility Center, New York University, New York, NY; ART Reproductive Center, Beverly Hills, CA; Fertility Centers of Illinois, Highland Park, IL; LifeQuest Centre for Reproductive Medicine, Toronto, ON; Main Line Fertility and Reproductive Medicine, Bryn Mawr, PA; Reprogenetics, Livingston, NJ. OBJECTIVE: Array CGH can analyze all chromosomes for aneuploidy and may further reduce the risk of miscarriage in patients suffering from recurrent pregnancy loss (RPL). The objective of this study was to determine any beneficial effects of PGD by array CGH for RPL patients in comparison to their expected loss rate and to PGD by FISH results. DESIGN: Multicenter retrospective study. MATERIALS AND METHODS: The study included 177 cycles of couples with idiopathic RPL (defined as 3 or more miscarriages), average maternal age of 36.3 years, undergoing ART at multiple fertility centers. PGD was done using day-3 biopsy (n ¼ 143) or day-5 biopsy (n ¼ 34), followed by analysis by aCGH. Each PGD patient was matched with their expected loss rate for RPL patients (Brigham et al. 1999). An ongoing pregnancy was defined as past second trimester. RESULTS: In total 1517 embryos were analyzed, of which 33% were normal, 63% were abnormal and 4% none analyzable (no nucleus or no DNA amplification). Of the 176 cycles, pregnancy data was available for 154, of which 66 (43%) became pregnant with an implantation rate of 45% (95 sacs / 212 replaced embryos) and 62 cycles (40%) are ongoing past second trimester or delivered. We would expect a 34.9% miscarriage rate in this specific group of patients, but the miscarriage rate found was only 6% (4/66) (P<0.001). CONCLUSION: Previous PGD results using FISH showed that the miscarriage rate in idiopathic RPL patients was significantly reduced from 26% to 10% in patients younger than 35, and from 39% to13% in older patients. Current PGD results with aCGH indicate a significant decrease in the miscarriage rate of idiopathic RPL patients and high pregnancy rates for the current average maternal age of 36. These data suggest that aCGH further improves the results previously obtained by FISH. Furthermore, this confirms that idiopathic recurrent miscarriage is mostly caused by embryonic chromosome abnormalities.
O-77 Monday, October 17, 2011 06:00 PM REPORT ON FIRST 54 PATIENTS UNDERGOING PREIMPLANTATION GENETIC DIAGNOSIS FOR GENETIC DISORDERS TOGETHER WITH 24 CHROMOSOME ANEUPLOIDY SCREENING USING MICROARRAYS. M. Rabinowitz, M. Hill, D. Potter, N. Wemmer, J. Keller, G. Gemelos. Gene Security Network, Redwood City, CA; HRC Fertility, Laguna Hills, CA. OBJECTIVE: To evaluate the clinical benefit of informatics-based SNP (single nucleotide polymorphism) microarray technology for preimplantation genetic diagnosis (PGD) for single gene (SG) disorders in parallel with 24-chromosome aneuploidy in at-risk couples. Aneuploidy PGD is usually performed separately from SG PGD. Current methods for SG PGD typically sequence the disease locus and nearby markers which can result in misdiagnosis of 1-2%. Efficiency and reliability can be improved by using SNP microarrays and Parental Support informatics to phase the parent homologs and infer recombination sites. DESIGN: Prospective non-randomized trial. MATERIALS AND METHODS: Any disease mutation was eligible for study inclusion. Referrals were accepted for testing following verification
FERTILITY & STERILITYÒ
that the mutation was disease-causing and there was adequate SNP microarray coverage at the gene loci. Embryo biopsy samples were shipped on dry ice to a laboratory for testing. All embryonic samples were compared to parent samples across multiple loci to rule out DNA contamination. RESULTS: 54 IVF cycles with 447 embryos were successfully tested for >30 mutations including triplet repeats, small in/dels, large deletions, and single nucleotide substitutions. 68% had a transfer on Day 5/6 with embryos testing unaffected by the disease and euploid across all 24 chromosomes. 71% had initial positive beta hcg and 55% had clinical pregnancy by positive fetal heart tone. All prenatal diagnosis and live birth results to date confirm the original PGD results. 15% of embryo samples were aneuploid on the disease-gene chromosome. On average there were 35 informative markers near the mutation site and confidence of genotype calls typically exceeded 99.9%. CONCLUSION: Reliable SG PGD in parallel with 24-chromosome aneuploidy screening can be performed with informatics enhanced SNP microarrays using a single cell. Preliminary results show highly accurate SG calls and 55% clinical pregnancy rates. Supported by: National Institutes of Health
PROCEDURES AND TECHNIQUES-CLINICAL: ART
O-78 Monday, October 17, 2011 04:15 PM THE VALUE OF FALLOPIAN TUBE SPERM PERFUSION IN THE MANAGEMENT OF MALE FACTOR INFERTILITY: A RANDOMIZED CONTROLLED TRIAL. W. M. El-Khayat, A. N. El-Mazny, N. F. Abou-salem, A. H. Moafy. Obstetrics & Gynecology, Faculty of Medicine, Cairo University, Cairo, Egypt; Clinical & Chemical Pathology, Faculty of Medicine, Cairo University, Cairo, Egypt. OBJECTIVE: To investigate whether fallopian tube sperm perfusion (FSP) improves pregnancy rates over standard intrauterine insemination (IUI) in male factor infertility. DESIGN: Randomized controlled trial. MATERIALS AND METHODS: The study population consisted of 120 couples allocated into two equal groups: group I (n ¼ 60): FSP with 4 ml of inseminate; and group II (n ¼ 60): IUI with 0.5 ml of inseminate. The study was done at Cairo University. All women underwent the same mild controlled ovarian stimulation protocol. Clomiphene citrate (Clomid; Aventis pharma S.AE, Global Napi Pharmaceuticals, Cairo, Egypt) 100 mg / day were administered orally from day 3 to 7 of the cycle. HMG (Merional; IBSA, Lugano, Switzerland) 75 IU / day was administered intramuscularly from day 6 to 8. Follicullometry was performed on day 9 of the cycle & HMG dose was adjusted, if necessary. When 2 to 3 follicles R18 mm in diameter were present, HCG (Choriomon; IBSA, Lugano, Switzerland) 10,000 IU was administered intramuscularly. A single insemination was performed 36 hours after HCG administration. On the insemination day, women were allocated into two groups with a ratio of 1:1 using computer-generated random numbers: group I (FSP using pediatric Foley catheter) and group II (standard IUI). Data analysis was performed using the Statistical Package for the Social Sciences, v16.0 (SPSS, Chicago, IL, USA). RESULTS: There was no statistically significant difference regarding patient or cycle characteristics. The pregnancy rate was significantly higher (P ¼ 0.0369) in group I (16/ 60; 26.7%) compared with group II (7/60; 11.7%). One case of twin pregnancy occurred in each group. Two abortions occurred in group I and one in group II, while one case of ectopic pregnancy occurred in group I. CONCLUSION: The FSP is a simple and effective technique in the management of mild or moderate male infertility.
O-79 Monday, October 17, 2011 04:30 PM EFFECT OF ELECTIVE SINGLE EMBRYO TRANSFER ON MULTIPLE PREGNANCY AND OVERALL PREGNANCY RATES AT THE OXFORD FERTILITY UNIT. N. Vulliemoz, A. Itani, T. Child, K. Turner, E. McVeigh. Oxford Fertility Unit, Oxford, Oxfordshire, United Kingdom. OBJECTIVE: Multiple birth is the most important complication of IVF/ ICSI. There is an increased risk of premature delivery and mortality. The
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O-73 Monday, October 17, 2011 05:00 PM HALVING THE MULTIPLE PREGNANCY RATE IN A PREIMPLANTATION GENETIC DIAGNOSIS PROGRAM BY ADOPTING A NEW BLASTOCYST FREEZING POLICY. L. Pepas, V. Bolton, Y. Khalaf, T. El-Toukhy. Asssisted Conception Unit, Guy’s and St Thomas’ Hospitals Trust, London, United Kingdom. OBJECTIVE: The implications of multiple pregnancy in patients with inherited genetic disorders are significant. Elective single embryo transfer (eSET) has been recommended for preimplantation genetic diagnosis (PGD) for inherited genetic disorders to reduce multiple pregnancy rates. As the safety and success of cryo-thawed PGD cycles has been demonstrated, we sought to evaluate effect of a stricter SET policy on the outcomes of our PGD programme. DESIGN: Retrospective study. MATERIALS AND METHODS: Since January 2006, patients who had two or more blastocysts suitable for transfer after biopsy and genetic analysis on Day 3 were offered eSET and cryopreservation of surplus expanded blastocyst(s). To increase the uptake of eSET further, the policy was changed in June 2009 to include cryopreservation of early blastocysts too. We compared the outcomes of 424 fresh PGD cycles performed between January 2006 and May 2009 prior to this change; to 279 cycles performed between June 2009 and December 2010. RESULTS: Embryo transfer was achieved in 71% and 73% of cycles in each group respectively (P¼0.68). There was no difference in mean age (33.6 3.9 and 32.8 4.1) and FSH values (6.8 2.1 and 6.7 2.1) between the two groups. Our elective SET rate increased from 44% to 61% (P¼0.0006) and blastocyst freezing increased from 28% to 43% (P¼0.0001). The mean number of embryos transferred decreased from 1.35 0.6 to 0.92 0.7 (P¼0.0001). Per cycle reaching embryo transfer, the pregnancy rate increased from 47% to 57% (P¼0.0230), the clinical pregnancy rate increased from 39% to 49% (P¼0.0274) and the ongoing pregnancy/live birth rate increased from 34% to 43% (P¼0.0305). The multiple pregnancy rate per clinical pregnancy halved from 13% (n ¼ 15) to 6% (n ¼ 7) (P¼0.117). CONCLUSION: Through adoption of a stricter elective SET policy, encouraged by the relaxation of criteria for blastocyst freezing, we halved our multiple pregnancy rate without compromising PGD treatment outcome. Supported by: This project was funded internally.
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Abstracts
O-74 Monday, October 17, 2011 05:15 PM FISH REANALYSIS OF INNER CELL MASS AND TROPHECTODERM SAMPLES OF PREVIOUSLY ARRAY-CGH SCREENED BLASTOCYSTS REVEALS HIGH ACCURACY OF DIAGNOSIS AND NO SIGN OF MOSAICISM OR PREFERENTIAL ALLOCATION. A. Capalbo, G. Wright, L. Themaat, T. Elliott, L. Rienzi, Z. P. Nagy. GENERA, Reproductive Medicine, Rome, Italy; Reproductive Biology Associates, Atlanta, GA. OBJECTIVE: To study the blastocyst genetic constitution (possible chromosomal mosaicism between ICM and TE and within TE), after isolating ICM by a novel method.and to evaluate array-CGH accuracy against FISH testing DESIGN: Cross-sectional study. MATERIALS AND METHODS: 32 blastocysts diagnosed as genetically abnormal by array-CGH were donated to research. ICM was isolated from the trophectoderm (TE) using a novel mechanical separation which was confirmed by KRT18 immunostaining. 24 of the 32 embryo had single or double aneuploidy and 8 aneusomic for unbalanced product of translocation or for long single arm deletions/duplications but with normal ploidy (by aCGH). After biopsy, individual samples were fixed by Carnoy method and analyzed with conventional 9-chromosomes FISH and locus specific probes. RESULTS: We reanalyzed 32 ICM and 96 TE samples (3 separate groups of cells from each TE of the same embryo). Looking at the single chromosome level, 28 out of 28 aneuploidies diagnosed by a-CGH were confirmed by FISH reanalysis both in ICM and TE samples resulting in a analytical sensitivity of 100%(95%CI 90.2-100). For almost all embryos, the reported aneuploidies were constitutive. As regard to chromosomes diagnosed as disomic, only 2 out of 248 (specificity 99.1 95%CI 97.4-99.8) were misdiagnosed by a-CGH, a trisomy 13 in an unbalanced embryo from reciprocal translocation carrier observed in a constitutive state and a mosaic nullisomy/monosomy 16 in a double aneuploid embryo. Consequently, only one out of 32 embryos was misdiagnosed as false negative. Only in one case we observed a confined trisomy 17 in all nuclei of a TE sample from a trisomic embryo for chromosome 13. In all other embryos ICM and TE samples were concordant and confirmed the original a-CGH diagnosis. CONCLUSION: Our data show a very high analytical accuracy of a-CGH to diagnose embryo chromosomal constitution. Concordance between ICM and TE was reported for all embryos analyzed, showing no sign of mosaicism or preferential allocation at blastocyst stage.
O-75 Monday, October 17, 2011 05:30 PM COMPARISON OF DAY-3 AND DAY-5 ARRAY-CGH DIAGNOSIS FOR 24 CHROMOSOME ANEUPLOIDY SCREENING IN TERMS OF ACCURACY. P. Mir, L. Rodrigo, E. Mateu, A. Cervero, J. Martın, C. Rubio. PGD Unit, IVI-Valencia, Valencia, Comunitat Valenciana, Spain. OBJECTIVE: To describe the clinical outcome on day-3 single cell arrayCGH (aCGH) and to compare day-3 and day-5 aCGH diagnosis DESIGN: Prospective study. MATERIALS AND METHODS: Day-3 aCGH diagnosis was performed on 24 patients. Indications for Preimplantation Genetic Screening (PGS) were: recurrent miscarriage, repetitive implantation failure, severe male factor and advanced maternal age. Mean female age was 38.8 2.9 years. A single cell from 122 day-3 embryos was amplified (SureplexÒ, BlueGnome, Cambridge, UK). DNA amplification, labelling, hybridisation, washing, scanning and data analysis (24sureÒplatform, BlueGnome) was performed in a 24-hour protocol. On day-5, 39 chromosomally abnormal embryos were reanalysed by FISH using probes for the altered chromosomes (Vysis Inc., Downers Grove, IL,USA). Moreover in 14 abnormal but good developing blastocysts, aCGH was performed on throphoectoderm biopsy and the remaining cells of the embryo were analysed by FISH. RESULTS: A successful amplification was obtained in 117 blastomeres (95.9%), being 74 embryos diagnosed as abnormal (63.2%); 30 of them (40.5%) had aneuploidies only for the chromosomes analysed in our FISH program (chromosomes 13, 15, 16, 17, 18, 21, 22, X, Y); 29 (39.2%) had aneuploidies also for other chromosomes; and 15 (20.3%) had aneuploidies only for other chromosomes. In 20 cycles euploid embryos were transferred (83.3%), a clinical pregnancy was observed in 15 patients (62.5% per cycle; 75% per transference) with an implantation rate of 54.3%.
Vol. 96., No.3, Supplement, September 2011
FISH re-analysis on day-5 confirmed day-3 aCGH diagnosis in 38 embryos (97.4%). aCGH on trophectoderm biopsy showed 93.3% (14/15) of concordance with day-3 aCGH. The discordant was chaotic aneuploid by aCGH on day-3; euploid by aCGH on trophectoderm biopsy but chaotic aneuploid by FISH re-analysis. CONCLUSION: PGS using aCGH is suitable on single blastomeres with successful amplification efficiency and reliable results. In this study, day-5 trophectoderm biopsy does not improve the accuracy of the diagnosis compared to day-3.
O-76 Monday, October 17, 2011 05:45 PM SIGNIFICANT DECREASE IN MISCARRIAGES AFTER PREIMPLANTATION GENETIC DIAGNOSIS (PGD) FOR RECURRENT PREGNANCY LOSS USING ARRAY COMPARATIVE GENOME HYBRIDIZATION (Array CGH). J. Grifo, S. Ghadir, B. Kaplan, C. A. Laskin, M. Glassner, S. Munne. NYU Fertility Center, New York University, New York, NY; ART Reproductive Center, Beverly Hills, CA; Fertility Centers of Illinois, Highland Park, IL; LifeQuest Centre for Reproductive Medicine, Toronto, ON; Main Line Fertility and Reproductive Medicine, Bryn Mawr, PA; Reprogenetics, Livingston, NJ. OBJECTIVE: Array CGH can analyze all chromosomes for aneuploidy and may further reduce the risk of miscarriage in patients suffering from recurrent pregnancy loss (RPL). The objective of this study was to determine any beneficial effects of PGD by array CGH for RPL patients in comparison to their expected loss rate and to PGD by FISH results. DESIGN: Multicenter retrospective study. MATERIALS AND METHODS: The study included 177 cycles of couples with idiopathic RPL (defined as 3 or more miscarriages), average maternal age of 36.3 years, undergoing ART at multiple fertility centers. PGD was done using day-3 biopsy (n ¼ 143) or day-5 biopsy (n ¼ 34), followed by analysis by aCGH. Each PGD patient was matched with their expected loss rate for RPL patients (Brigham et al. 1999). An ongoing pregnancy was defined as past second trimester. RESULTS: In total 1517 embryos were analyzed, of which 33% were normal, 63% were abnormal and 4% none analyzable (no nucleus or no DNA amplification). Of the 176 cycles, pregnancy data was available for 154, of which 66 (43%) became pregnant with an implantation rate of 45% (95 sacs / 212 replaced embryos) and 62 cycles (40%) are ongoing past second trimester or delivered. We would expect a 34.9% miscarriage rate in this specific group of patients, but the miscarriage rate found was only 6% (4/66) (P<0.001). CONCLUSION: Previous PGD results using FISH showed that the miscarriage rate in idiopathic RPL patients was significantly reduced from 26% to 10% in patients younger than 35, and from 39% to13% in older patients. Current PGD results with aCGH indicate a significant decrease in the miscarriage rate of idiopathic RPL patients and high pregnancy rates for the current average maternal age of 36. These data suggest that aCGH further improves the results previously obtained by FISH. Furthermore, this confirms that idiopathic recurrent miscarriage is mostly caused by embryonic chromosome abnormalities.
O-77 Monday, October 17, 2011 06:00 PM REPORT ON FIRST 54 PATIENTS UNDERGOING PREIMPLANTATION GENETIC DIAGNOSIS FOR GENETIC DISORDERS TOGETHER WITH 24 CHROMOSOME ANEUPLOIDY SCREENING USING MICROARRAYS. M. Rabinowitz, M. Hill, D. Potter, N. Wemmer, J. Keller, G. Gemelos. Gene Security Network, Redwood City, CA; HRC Fertility, Laguna Hills, CA. OBJECTIVE: To evaluate the clinical benefit of informatics-based SNP (single nucleotide polymorphism) microarray technology for preimplantation genetic diagnosis (PGD) for single gene (SG) disorders in parallel with 24-chromosome aneuploidy in at-risk couples. Aneuploidy PGD is usually performed separately from SG PGD. Current methods for SG PGD typically sequence the disease locus and nearby markers which can result in misdiagnosis of 1-2%. Efficiency and reliability can be improved by using SNP microarrays and Parental Support informatics to phase the parent homologs and infer recombination sites. DESIGN: Prospective non-randomized trial. MATERIALS AND METHODS: Any disease mutation was eligible for study inclusion. Referrals were accepted for testing following verification
FERTILITY & STERILITYÒ
that the mutation was disease-causing and there was adequate SNP microarray coverage at the gene loci. Embryo biopsy samples were shipped on dry ice to a laboratory for testing. All embryonic samples were compared to parent samples across multiple loci to rule out DNA contamination. RESULTS: 54 IVF cycles with 447 embryos were successfully tested for >30 mutations including triplet repeats, small in/dels, large deletions, and single nucleotide substitutions. 68% had a transfer on Day 5/6 with embryos testing unaffected by the disease and euploid across all 24 chromosomes. 71% had initial positive beta hcg and 55% had clinical pregnancy by positive fetal heart tone. All prenatal diagnosis and live birth results to date confirm the original PGD results. 15% of embryo samples were aneuploid on the disease-gene chromosome. On average there were 35 informative markers near the mutation site and confidence of genotype calls typically exceeded 99.9%. CONCLUSION: Reliable SG PGD in parallel with 24-chromosome aneuploidy screening can be performed with informatics enhanced SNP microarrays using a single cell. Preliminary results show highly accurate SG calls and 55% clinical pregnancy rates. Supported by: National Institutes of Health
PROCEDURES AND TECHNIQUES-CLINICAL: ART
O-78 Monday, October 17, 2011 04:15 PM THE VALUE OF FALLOPIAN TUBE SPERM PERFUSION IN THE MANAGEMENT OF MALE FACTOR INFERTILITY: A RANDOMIZED CONTROLLED TRIAL. W. M. El-Khayat, A. N. El-Mazny, N. F. Abou-salem, A. H. Moafy. Obstetrics & Gynecology, Faculty of Medicine, Cairo University, Cairo, Egypt; Clinical & Chemical Pathology, Faculty of Medicine, Cairo University, Cairo, Egypt. OBJECTIVE: To investigate whether fallopian tube sperm perfusion (FSP) improves pregnancy rates over standard intrauterine insemination (IUI) in male factor infertility. DESIGN: Randomized controlled trial. MATERIALS AND METHODS: The study population consisted of 120 couples allocated into two equal groups: group I (n ¼ 60): FSP with 4 ml of inseminate; and group II (n ¼ 60): IUI with 0.5 ml of inseminate. The study was done at Cairo University. All women underwent the same mild controlled ovarian stimulation protocol. Clomiphene citrate (Clomid; Aventis pharma S.AE, Global Napi Pharmaceuticals, Cairo, Egypt) 100 mg / day were administered orally from day 3 to 7 of the cycle. HMG (Merional; IBSA, Lugano, Switzerland) 75 IU / day was administered intramuscularly from day 6 to 8. Follicullometry was performed on day 9 of the cycle & HMG dose was adjusted, if necessary. When 2 to 3 follicles R18 mm in diameter were present, HCG (Choriomon; IBSA, Lugano, Switzerland) 10,000 IU was administered intramuscularly. A single insemination was performed 36 hours after HCG administration. On the insemination day, women were allocated into two groups with a ratio of 1:1 using computer-generated random numbers: group I (FSP using pediatric Foley catheter) and group II (standard IUI). Data analysis was performed using the Statistical Package for the Social Sciences, v16.0 (SPSS, Chicago, IL, USA). RESULTS: There was no statistically significant difference regarding patient or cycle characteristics. The pregnancy rate was significantly higher (P ¼ 0.0369) in group I (16/ 60; 26.7%) compared with group II (7/60; 11.7%). One case of twin pregnancy occurred in each group. Two abortions occurred in group I and one in group II, while one case of ectopic pregnancy occurred in group I. CONCLUSION: The FSP is a simple and effective technique in the management of mild or moderate male infertility.
O-79 Monday, October 17, 2011 04:30 PM EFFECT OF ELECTIVE SINGLE EMBRYO TRANSFER ON MULTIPLE PREGNANCY AND OVERALL PREGNANCY RATES AT THE OXFORD FERTILITY UNIT. N. Vulliemoz, A. Itani, T. Child, K. Turner, E. McVeigh. Oxford Fertility Unit, Oxford, Oxfordshire, United Kingdom. OBJECTIVE: Multiple birth is the most important complication of IVF/ ICSI. There is an increased risk of premature delivery and mortality. The
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