DESIGN: Prospective randomized controlled trial (RCT) started in June 2012with preliminary outcome evaluated in December 2013. Patients were allocatedthrough computer-generated randomization into two groups: conventional blas-tocyst transfer or CCS cycle. Sample size calculated for the endpoint of ongoingpregnancy rates per cycle and delivery rates was 120 patients per armwith ovumpick-up (a 5%, b 20%). ClinicalTrials.gov NCT01571076.

MATERIALS AND METHODS: Inclusi�on criteria were: sperm concen-tration<2 million/mL; women<38 years;<2 implantation failures;<2 pre-vious miscarriages, normal karyotypes and >5 MII oocytes retrieved.Embryo culture and fresh blastocyst transfer was performed in the non-inter-vention arm. Cleavage stage embryo biopsy and array CGH were performedin the intervention arm with fresh blastocyst transfer of euploid embryos. In-dividual blastomeres underwent whole genome amplification using Sureplex(BlueGnome Ltd., UK). Amplified DNA and reference DNA were labeledand co-hybridized in 24sure arrays (BlueGnome Ltd., UK). After washing,slides were scanned and analyzed by BlueFuse Multi software. Main out-comes were ongoing pregnancy rates per transfer and per cycle Stastisticalcomparisons were performed using Fisher�sexact test.

RESULTS: In the group of blastocyst transfer, 35 cycles were completedwith 33 transfers and 15 ongoing pregnancies (45.4 ongoing pregnancy rateper transfer and 42.8 per cycle). In the CCS group, 33 cycles were performedwith 31 transfers and 22 ongoing pregnancies (71.0 ongoing pregnancy rateper transfer and 66.7 per cycle). One-side Chi-square test showed significantdifferences for ongoing pregnancy rates per transfer (p¼0.0345) and per cy-cle (p<0.0417). In the blastocyst transfer group, miscarriage rate was 28.6,whereas no miscarriage was observed in the CCS group.

CONCLUSION: Embryo selection based in 24-chromosome aneuploidyscreeningcouldbe consideredas avaluable clinical tool to assess embryoviabilityin severe male factor patients, since significantly improved clinical outcome.

Supported by: BlueGnome (Illumina), IVI, IVIOMICS.

O-71 Monday, October 20, 2014 04:30 PM

WHOLE EXOME SEQUENCING OF EMBRYO BIOPSIESREVEALS CLINICALLY-SIGNIFICANT DE NOVOMUTATIONS. N. Steuerwald,a R. Durrett,b J. Parsons,a A. Hamilton,a

M. Kontanstinidis,c F. Licciardi,d S. Munne.c aCarolinas Medical Center, Char-lotte, NC; bRecombine, New York, NY; cReprogenetics, Livingston, NJ; dNewYourk University Fertility Center, New York University, New York, NY.

OBJECTIVE: Current PGD techniques can detect any inheritable genedefect identified in the parents by carrier screening. However, de novomutations occurring in the germ line, some linked to autism and other syn-dromes, cannot be detected unless through whole genome or targetedsequencing. The objective of this study was to detect pathogenic de novomu-tations in blastocyst biopsies by sequencing the whole exome of the embryo.

DESIGN: Whole exome analysis by Next Generation Sequencing (NGS)was performed on embryo biopsies and compared to the parents wholeexome to rule out inherited mutations and identify de novo ones.

MATERIALS AND METHODS: An embryo-parent trio was profiled formutations by whole exome sequencing. DNA isolated from a blastocyst bi-opsy was subjected towhole genome amplification using a multiple displace-ment amplification. Sequencing libraries were enriched for exonic regions bytargeted multiplex PCR. These PCR products were sequenced on an Ion Pro-tonTM Sequencer with Ion PI Chips. NGS data analysis platform from SevenBridges Genomics utilizing GATK was used to align reads, recalibrate basequalities, and call variants on the sequencing data. Variant calls were thencontrasted between the parent and embryo data, and variants unique to theembryo were further analyzed.

RESULTS: 73,774 variants were identified of which 538were de novo var-iants discovered only in the embryo biopsy. Approximately 52% of these var-iants were synonymous and the other 48% were missense. Most de novomutations were identified to have benign biological consequences, some(including 16 in splice junction regions and 1 stop-loss mutation) have un-known significance, and two de novo variants were marked with clinical sig-nificance in the TOR1A and TYR genes, which, through loss of function, cancause Torsion Dystonia and Albinism, respectively.

CONCLUSION: Trio-based exome sequencing can yield importantclinically-relevant information regarding preimplantation embryos notfound in the parents. While sequencing data from amplified DNA sam-ples may be noisy, high-threshold quality filtering can be used to main-tain high confidence in de novo variants. While genetic counseling isin order, this type of analysis could be utilized in routine PGS treatmentto gain a broader understanding of the genetics of the embryos availablefor replacement.

FERTILITY & STERILITY�

O-72 Monday, October 20, 2014 04:45 PM

NGS VERSUS CGH FOR THE DETECTION OFWHOLE CHROMO-SOME AND SEGMENTAL ANEUPLOIDIES IN HUMANBLASTOCYSTS. N. Al-Asmar,a M. Vera,b C. E. Michel,c

C. Cinnioglu,a D. Blesa,b C. Simon,b,d,e,f C. Rubio.b,d aIviGen, Miami, FL;bIVIOMICS, Paterna, Valencia, Spain; cIllumina, Fulbourn, Cambridge, UnitedKingdom; dFundaci�on Instituto Valenciano de Infertilidad, Incliva, Valencia,Spain; eDepartment of Obstetrics and Gynecology, Stanford University, CA.

OBJECTIVE: To evaluate the capability of next-generation sequencing(NGS) to detect whole chromosome and segmental aneuploidies in trophecto-derm biopsies and to define the concordance rate with the results obtained withthe current platform of array comparative genomic hybridization (aCGH).DESIGN: Validation study of aNGSplatformusing amplifiedDNA from tro-

phectoderm biopsies in which whole or partial chromosome aneuploidies werepreviously detected by aCGH using comprehensive chromosome screening.MATERIALS AND METHODS: Trophectoderm samples from each

embryo underwent whole genome amplification using the Sureplex Kit (Blue-Gnome, UK). For aCGH, samples and reference DNAwere labeled and co-hy-bridized in 24sure arrays. After washing, slides were scanned and analyzed byBlueFuse Multi software (BlueGnome, UK). For NGS, 1ng dsDNAwas tag-mented and amplified with the addition of barcodes. Each library was normal-ized before pooling and loaded into a MiSeq instrument (Illumina., UK).RESULTS: Aneuploidies were classified into whole chromosome and

segmental events according to aCGH profiles. A total of 120 aneuploidiesfrom 64 embryos were selected. For whole chromosome aneuploidies(n¼45), the concordance rate was 100%. Seventy-five segmental eventswere also identified in the same embryos. The chromosome segment lengthranged between 9.0 and 187.8Mb. The concordance rate in this casewas 96%(72/75). Segmental aneuploidies were classified as control (n¼28), when onemember of the couple was a carrier of a reciprocal translocation, or studycases (n¼47) when parental karyotypes were described as normal. Theconcordance rate for the control cases was 100% (28/28) and 93.6% forthe study cases (44/47). Chi-square test showed no significant differencesin the concordance rate for segmental aneuploidies between the controland study group (p¼0.45, 95% CI, 80% statistical power).CONCLUSION: NGS allows the complete detection of whole chromo-

some aneuploidies in human blastocyst with the same efficiency as aCGH.For the detection of segmental aneuploidies a high concordance rate wasfound between the two platforms. Studies like this are essential for the devel-opment of an appropriate software that allows the efficient translation ofNGS into the CCS programs.

O-73 Monday, October 20, 2014 05:00 PM

CLINICAL EXPERIENCE WITH KARYOMAPPING FOR PREIM-PLANTATION GENETIC DIAGNOSIS (PGD) OF SINGLE GENEDISORDERS. R. Prates,a M. Konstantinidis,a N.-N. Goodall,a

J. Fischer,a W. Hummel,b J. Grifo,c C. Laskin,d J. Hesla,e

D. Tourgeman,f D. Wells,g S. Munne.a aReprogenetics, Livingston, NJ;bSan Diego Fert Center, San Diego, CA; cNYU Fert Center, New York,NY; dLifeQuest Centre Reprod Medicine, Toronto, ON, Canada; eOregonReprod Medicine, Portland, OR; fHRC Fert., Encino, CA; gReprogeneticsUK, Oxford, Oxfordshire, United Kingdom.

OBJECTIVE: To clinically assess the efficacy of Karyomapping (Kmap)for PGD of single gene disorders.DESIGN: A total of 43 clinical cases for PGD of single gene disorders

were carried out using Kmap. For the first 23 of the cases, a conventionalPGD test [short tandem repeat (STR) linkage analysis and direct mutationdetection] was carried out in parallel with Kmap. For the rest of the cases,Kmap was performed with direct mutation detection alone.MATERIALS AND METHODS: 181 blastocysts were biopsied and the

obtained cells amplified with multiple displacement amplification. Separatealiquots of product were used to carry out the Kmap protocol and the vali-dated STR and direct mutation tests.RESULTS: Successful amplification was detected in 91.7% (166/181) of

the biopsied samples. 86 embryos were given a diagnosis by Kmap and con-ventional PGD testing. Results obtained were in complete concordance(100%) between the two tests. Kmap was able to provide diagnosis on anadditional 5 embryos that successfully amplified but received an inconclusivediagnosis using the conventional test due to allele dropout (ADO) or recom-bination. For the rest of the cases where Kmapwas used in parallel with directmutation detection, results agreed between the two methods for 74/75(98.7%) embryos (discordance observed was due to ADO affecting the

e25

mutation site). Currently, there are 2 ongoing clinical pregnancies which re-sulted from transfer of 3 embryos diagnosed as unaffected using Kmap.

CONCLUSION: Kmap utilize a universal protocol to performPGD for SGD.Asa result it reduces significantly the time that patients have towait to start their invitro fertilization cycle. As shown by this study, Kmap can offer a more compre-hensive assessment of the region of interest than conventional STR analysis, al-lowing for more embryos to receive diagnosis. Data from Kmap demonstratesthat the method is highly accurate. Nonetheless, inclusion of a direct mutationtest may be valuable in cases where recombination occurs close to the gene of in-terest, when the mutant gene is in a region with reduced SNP coverage (e.g. telo-meric regions) or when offering PGD for consanguineous couples.

O-74 Monday, October 20, 2014 05:15 PM

ANEMBRYOCOHORTWHICHCONTAINSALLANEUPLOIDEM-BRYOS IS NOT INDICATIVE OF FUTURE EMBRYO COHORTANEUPLOIDY. J. M. Franasiak,a,b J. Horne,b K. H. Hong,a,b

M. D. Werner,a,b R. T. Scott, Jr.,a,b aRWJ Medical School, Rutgers Univer-sity, New Brunswick, NJ; bRMA of New Jersey, Basking Ridge, NJ.

OBJECTIVE: Use of comprehensive chromosome screening (CCS) hasenhanced embryo selection. However, some patients will have 100% of theircohort return as aneuploid. This varies from 5% in women in their late 20s to55% in women >45. Beyond the initial disappointment, patients and clini-cians are left with the difficult decision about whether or not to attemptanother IVF cycle. Is a cohort which is 100% aneuploid indicative of futureaneuploidy rates? The data presented here seek to answer those questions andassist in patient counseling and clinical decision making.

DESIGN: Retrospective cohort study.MATERIALS AND METHODS: All patients undergoing their first IVF

cycle with CCS who had complete embryonic aneuploidy were selectedfor inclusion. This cohort was then followed through subsequent IVF/CCScycles to determine how often the embryo cohort resulted in all aneuploidembryos. The total number of euploid and aneuploid embryos was recordedfor each patient as well as the percent of complete embryonic aneuploidy persubsequent IVF/CCS cycle.

RESULTS: In 2012-2014, 316 patients had complete embryonic aneuploidyand 128 of these patients pursued a subsequent cycle. Of all embryos screenedin a subsequent cycle, 43.8% were euploid. For all patients undergoing a sub-sequent cycle, 59.4%eventuallyhad at least 1 euploidembryo.Of those patientsthat had complete embryonic aneuploidy in the 2nd consecutive cycle, 20 pur-sued an additional cycle for an overall embryonic euploid rate of 39.9%.Withinthis group, 60% had at least 1 euploid embryo in a subsequent cycle. Of thosewith 3 consecutive cycles with complete embryonic aneuploidy, only 3 patientscompleted a subsequent cycle with an overall euploid rate of 20%.

Patients with an all aneuploid embryo cohort are likely to have a euploid

embryo in subsequent cycles.

SubsequentCycle #

Patients with R1Euploid Embryo

n (%)Euploid Embryos

n (%) Mean Age

1 76/128 (59.4) 147/336 (43.8) 39.72 12/20 (60.0) 24/56 (42.9) 39.93 2/3 (66.6) 2/10 (20.0) 41.9

Correlation between Paternal Age and Euploidy Rate

SARTMaternalAgeCategory(years) n

AveragePaternalAge

(ranges)

PearsonCorrelationCoefficientbetween

Paternal Age andEuploidy Rate

Pearson CorrelationCoefficient betweenPateranl Age and

%2PNs thatmade it tobiopsy

<35 148 36.8 (22-71) 0.1 (p¼0.22) -0.02 (p¼0.8)35-37 115 38.9 (29-59) -0.131 (p¼0.16) 0.104 (p¼0.27)38-40 181 42 (31-82) -0.021 (p¼0.78) -0.102 (p¼0.17)41-42 135 43.6 (33-71) -0.045 (p¼0.6) 0.02 (p¼0.82)>42 79 44.8 (34-68) 0.135 (p¼0.24) 0.038 (p¼0.73)

CONCLUSION: In patients who undergo IVF/CCS which results in an allaneuploid embryo cohort, they are likely to have a euploid embryo with sub-sequent cycles. These data are limited by the drop-out rate due to patientswho do not have a euploid embryo and do not cycle again. However, it clearlydemonstrates that presence of an all aneuploid embryo cohort should not pre-vent a repeat cycle of IVF/CCS.

O-75 Monday, October 20, 2014 05:30 PM

PUBLIC PERSPECTIVES ON PREIMPLANTATION GENETICDIAGNOSIS (PGD) FROM A NATIONALLY REPRESENTATIVESAMPLE. W. Winkelman,a S. Missmer,b D. Myers,b E. Ginsburg.baUniversity of California, San Francisco, SF, CA; bBrigham and Women’sHospital, Boston, MA.

OBJECTIVE: Approximately 5% of in vitro fertilization procedures uti-lize PGD. Public perspectives of PGD may inform policies by professionalorganizations and fertility centers.

e26 ASRM Abstracts

DESIGN: Online survey through Harris Interactive Services of a represen-tative sample of U.S. residents aged 18-75 years, literate in English and withinternet access.MATERIALS AND METHODS: Responses to questions about PGD in

various clinical scenarios were collected from Nov to Dec 2013. Multivari-able logistic regression was applied to calculate odds ratios (OR) and 95%Wald confidence intervals (CI).RESULTS: Of 1006 respondents, a majority Supported PGD for diseases

fatal early in life or those causing lifelong disability (72.9% and 66.7%,respectively); only 48.0% Supported PGD for diseases that manifest late inlife. Respondents were more supportive of PGD for genetic diseases ifthey were aware of PGD prior to the survey (OR¼1.64; CI¼1.13-2.39).Among the scenarios for genetically-based trait selection, 21.1% SupportedPGD for sex selection, 14.6% for physical traits, 18.9% for personality traits,and 13.4% for sexual orientation. Compared to women, men were two tothree fold more supportive of PGD for sex selection (OR¼1.65; CI¼1.20-2.78), physical traits (OR¼2.38; CI¼1.60-3.48) personality traits(OR¼2.31; CI¼1.64-3.26) and sexual orientation (OR¼2.73; CI¼1.82-4.09). Compared to Caucasians, Asians were four times more supportiveof PGD for sex selection (OR¼3.87; CI¼1.71-8.78) and African Americanswere less opposed to PGD for sex selection (OR¼0.52; CI¼0.34-0.80), phys-ical traits (OR¼0.51; CI¼0.34-0.78), or personality traits (OR¼0.53;CI¼0.35-0.81). Those with a graduate degree were more opposed to PGDfor sex selection (OR¼2.36; CI¼1.49-3.74), physical traits (OR¼ 2.06;CI¼1.28-3.31) and personality traits (OR¼2.72; CI¼1.70-4.35) comparedto those without a college degree. Respondents who Supported PGD to iden-tify genetic diseases were not more likely to support PGD for genetic trait se-lection (chi-square 2-sided p-value<0.001).CONCLUSION: In a nationally representative sample, a majority support

PGD to identify diseases that present in childhood, but most did not supportPGD for diseases that manifest late in life. We noted significant variation inopinions by the gender, race, and education. There was more support amongthose with prior knowledge of PGD suggesting that education about PGDfosters favorable opinions. This study raises questions about public knowl-edge and attitudes that may shape future research and policies.Supported by: The research was funded by a grant from Brigham and

Women’s Department of Obstetrics and Gynecology.

O-76 Monday, October 20, 2014 05:45 PM

INCREASINGPATERNALAGE IS NOTASSOCIATEDWITHANEU-PLOIDY IN PREIMPLANTATION EMBRYOS. S. Morin,a B. Hodes-Wertz,b J. Grifo.b aOb/Gyn, NYU Langone Medical Center, New York, NY;bNYU Fertility Center, NYU Langone Medical Center, New York, NY.

OBJECTIVE: To determine whether the incidence of aneuploidy as deter-mined by preimplantation genetic screening (PGS) is associated withincreasing paternal age.DESIGN: Retrospective cohort.MATERIALS AND METHODS: 658 PGS cycles between 10/2010 and 2/

2014 were evaluated. In all cases, PGS occurred after trophectoderm biopsyon day 5 or 6, and chromosomal evaluationwas performed by array comparativegenomic hybridization. Each PGS cyclewas categorized according to the SARTcriteria for maternal age (<35, 35-37, 38-40, 41-42, >42 years). Within eachSART group, a linear regression analysis was performed to evaluate whetherincreasing paternal age was associated with the percentage of euploid embryosin a given cohort. Within the<35 year old maternal group, euploidy rates werealso compared in men <30 years versus men >45 years by student’s t-test.RESULTS: There was no consistent correlation between increasing

paternal age and euploidy rate amongst the SART groups.

Vol. 102, No. 3, Supplement, September 2014