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1.
Ashley W. Tiegs Brooke Hodes-Wertz David H. McCulloh Santiago Munné James A. Grifo 《Journal of assisted reproduction and genetics》2016,33(7):893-897
Purpose
Preimplantation genetic screening (PGS) and diagnosis (PGD) with euploid embryo transfer is associated with improved implantation and live birth rates as compared to routine in vitro fertilization. However, misdiagnosis of the embryo is a potential risk. The purpose of this study was to investigate the clinical discrepant diagnosis rate associated with transfer of trophectoderm-biopsied blastocysts deemed to be euploid via array comparative genomic hybridization (aCGH).Methods
This is a retrospective cohort study including cycles utilizing PGS or PGD with trophectoderm biopsy, aCGH, and euploid embryo transfer at a large university-based fertility center with known birth outcomes from November 2010 through July 2014 (n?=?520).Results
There were 520 embryo transfers of 579 euploid embryos as designated by aCGH. Five discrepant diagnoses were identified. Error rate per embryo transfer cycle was 1.0 %, 0.9 % per embryo transferred, and 1.5 % per pregnancy with a sac. The live birth (LB) error rate was 0.7 % (both sex chromosome errors), and the spontaneous abortion (SAB) error rate was 17.6 % (3/17 products of conception tested, but could range from 3/42 to 7/42). No single gene disorders were mistakenly selected for in any known cases.Conclusions
Although aCGH has been shown to be a highly sensitive method of comprehensive chromosome screening, several possible sources of error still exist. While the overall error rate is low, these findings have implications for counseling couples that are contemplating PGS and PGD with aCGH.2.
Jonathan D. Kort Ruth B. Lathi Kathleen Brookfield Valerie L. Baker Qianying Zhao Barry R. Behr 《Journal of assisted reproduction and genetics》2015,32(6):925-930
Purpose
Studies have demonstrated high implantation rates after trophectoderm biopsy of day 5 expanded blastocysts. However, biopsy of cleavage stage embryos may adversely affect embryo development and implantation. No studies have assessed the utility of day 5 morulae and early blastocyst biopsy. This study sought to better understand these slower embryos’ aneuploidy rates and implantation potential.Methods
This was a retrospective review of all autologous IVF cycles utilizing PGS at a single academic infertility center.Results
The biopsy of day 5 morulae and early blastocysts provided 22 % additional euploid blastocysts available for fresh day 6 transfer compared to day 5 biopsy of only expanded blastocysts. Aneuploidy did correlate with embryo stage on day 5, even after controlling for maternal age, with 16 % of morulae and 35 % of blastocysts being euploid. The majority (83 %) of euploid morulae progressed to the blastocyst stage by day 6. Experience transferring slower developing embryos is limited, but preliminary pregnancy and implantation rates appear similar to euploid embryos biopsied as expanded blastocysts.Conclusions
The biopsy of all non-arrested embryos on day 5 provides genetic information for all blastocysts on day 6, increasing the pool of euploid blastocysts available for fresh transfer and avoiding the need to cryopreserve developmentally competent embryos without genetic information.3.
Haitao Wu Xiaoting Shen Lei Huang Yanhong Zeng Yumei Gao Lin Shao Baomin Lu Yiping Zhong Benyu Miao Yanwen Xu Yali Wang Yubin Li Luoxing Xiong Sijia Lu X. Sunney Xie Canquan Zhou 《Journal of assisted reproduction and genetics》2018,35(6):1071-1078
Purpose
This paper aims to investigate the feasibility of performing pre-implantation genetic diagnosis (PGD) and pre-implantation genetic screening (PGS) simultaneously by a universal strategy without the requirement of genotyping relevant affected family members or lengthy preliminary work on linkage analysis.Methods
By utilizing a universal Mutated Allele Revealed by Sequencing with Aneuploidy and Linkage Analyses (MARSALA) strategy based on low depth whole genome sequencing (~3x), not involving specific primers' design nor the enrichment of SNP markers for haplotype construction. Single-sperm cells and trephectoderm cells from in vitro fertilized embryos from a couple carrying HBB mutations were genotyped. Haplotypes of paternal alleles were constructed and investigated in embryos, and the chromosome copy number profiles were simultaneously analyzed.Results
The universal MARSALA strategy allows the selection of a euploid embryo free of disease mutations for in uterus transfer and successful pregnancy. A follow-up amniocentesis was performed at 17 weeks of gestation to confirm the PGD/PGS results.Conclusion
We present the first successful PGD procedure based on genotyping multiple single-sperm cells to obtain SNP linkage information. Our improved PGD/PGS procedure does not require genotyping the proband or relevant family members and therefore can be applicable to a wider population of patients when conducting PGD for monogenic disorders.4.
E. Rocafort M. Enciso A. Leza J. Sarasa J. Aizpurua 《Journal of assisted reproduction and genetics》2018,35(9):1573-1583
Purpose
We investigated if automated TLI selection may be a valuable strategy to identify those euploid embryos with the best chances of success.Methods
This is a unicentric and retrospective study involving 244 patients undergoing preimplantational genetic screening (PGS) cycles with autologous oocytes or oocyte donation (OD) with single euploid embryo transferred. We examined euploid embryos selected for transfer based on morphology evaluation alone (PGS-only; control group) or by assessment using an automated TLI system (Eeva?; PGS-TLI group).Results
In both, autologous oocytes and OD patients, significantly better implantation and clinical and ongoing pregnancy rates were obtained in the PGS-TLI group when euploid embryos with high implantation potential as predicted by the automated TLI System (Eeva?) were transferred compared with the PGS-only group. This improvement was also observed when only transfers of good morphological quality embryos were compared. TLI categories showed significant differences on blastocyst formation and euploidy rate.Conclusions
Automated TLI combined with PGS is a useful prognostic tool to identify euploid embryos with the highest potential for implantation and pregnancy. Further, these results provide evidence that a healthy pregnancy does not only depend upon normal chromosomal status.5.
S. Mateo F. Vidal M. Parriego I. Rodríguez V. Montalvo A. Veiga M. Boada 《Journal of assisted reproduction and genetics》2017,34(7):905-911
Purpose
The purpose of this study was to investigate the chromosomal constitution and the developmental potential of intracytoplasmic sperm injection (ICSI) deriving embryos displaying a single pronucleus at the zygote stage.Methods
Eighty-eight embryos from single pronucleus (1PN) two polar bodies (2PB) ICSI zygotes from 64 preimplantational genetic screening (PGS) cycles (October 2012–December 2014), were retrospectively analyzed. Zygotes were cultured in a time-lapse incubator. Embryo biopsy was performed on day 3 and genetic analysis approached by array comparative genomic hybridization.Results
Chromosomal analysis revealed that 17% (15/88) of embryos derived from 1PN 2PB zygotes were diagnosed as euploid. After blastomere biopsy at day 3, the blastocyst rate at day 5 was 3.4% (3/88). Only 2.3% (2/88) euploid blastocysts were obtained. In two couples and after counseling and patient agreement, the transfer of a euploid blastocyst from a 1PN 2PB ICSI zygote was performed resulting in the birth of a healthy child.Conclusions
These results open the possibility to consider embryos coming from 1PN 2PB ICSI zygotes for transfer when no other embryos from 2PN 2PB ICSI zygotes are available and if a PGS diagnosis of euploidy is obtained. Confirmation of biparental inheritance is strongly recommended.6.
Alexis P Melnick Robert Setton Logan D. Stone Nigel Pereira Kangpu Xu Zev Rosenwaks Steven D. Spandorfer 《Journal of assisted reproduction and genetics》2017,34(10):1325-1331
Purpose
The goal of this study was to compare pregnancy outcomes between natural frozen embryo transfer (FET) cycles in ovulatory women and programmed FET cycles in anovulatory women after undergoing in vitro fertilization with preimplantation genetic screening (IVF-PGS).Methods
This was a retrospective cohort study performed at an academic medical center. Patients undergoing single FET IVF-PGS cycles between October 2011 and December 2014 were included. Patients were stratified by type of endometrial replacement: programmed cycles with estrogen/progesterone replacement and natural cycles. IVF-PGS with 24-chromosome screening was performed on all included patients. Those patients with euploid embryos had single embryo transfer in a subsequent FET. The primary study outcome was live birth/ongoing pregnancy rate. Secondary outcomes included implantation, biochemical pregnancy, and miscarriage rates.Results
One hundred thirteen cycles met inclusion criteria: 65 natural cycles and 48 programmed cycles. The programmed FET group was younger (35.9 ± 4.5 vs. 37.5 ± 3.7, P = 0.03) and had a higher AMH (3.95 ± 4.2 vs. 2.37 ± 2.4, P = 0.045). The groups were similar for BMI, gravidity, parity, history of uterine surgery, and incidence of Asherman’s syndrome. There was also no difference in embryo grade at biopsy or transfer, and proportion of day 5 and day 6 transfers. Implantation rates were higher in the natural FET group (0.66 ± 0.48 vs. 0.44 ± 0.50, P = 0.02). There was no difference in the rates of biochemical pregnancy or miscarriage. After controlling for age, live birth/ongoing pregnancy rate was higher in natural FETs with an adjusted odds ratio of 2.68 (95% CI 1.22–5.87).Conclusions
Natural FET in ovulatory women after IVF-PGS is associated with increased implantation and live birth rates compared to programmed FET in anovulatory women. Further investigation is needed to determine whether these findings hold true in other patient cohorts.7.
Amy Kaing Lindsay L. Kroener Robyn Tassin Man Li Lian Liu Richard Buyalos Gary Hubert Mousa Shamonki 《Journal of assisted reproduction and genetics》2018,35(1):119-125
Purpose
The purpose of this study is to evaluate whether day of blastocyst development is associated with embryo chromosomal status as determined by high-density oligonucleotide microarray comparative genomic hybridization (aCGH).Methods
This is a retrospective cohort analysis, including women who underwent in vitro fertilization (IVF) with trophectoderm biopsy at a single private fertility center from January 2014 to December 2014. Repeat cycles were excluded. Cycles were assessed for percentage of blastocysts biopsied on days 5, 6, or 7 and rate of euploid embryos per cycle. Cycles were stratified by Society for Assisted Reproductive Technology (SART) age groups (< 35, 35–37, 38–40, 41–42, > 42) and by donor status.Results
A total of 388 IVF cycles and 2132 biopsied blastocysts were evaluated. The percentages of blastocysts biopsied on days 5, 6, and 7 were 62.5, 35.8, and 1.7%, respectively. Blastocyst euploid rates on days 5, 6, and 7 were 49.5, 36.5, and 32.9%, respectively. Earlier blastocyst development was associated with a significantly increased euploid rate (p < 0.0001). Younger maternal age (p < 0.0001) and higher number of blastocysts biopsied per patient (p = 0.0063) were both independently associated with greater percentage of euploidy.Conclusions
Earlier blastocyst development is independently associated with a higher likelihood of embryonic euploidy in both autologous and donor embryos. In non-biopsied embryos, these data support selection of day 5 blastocysts for transfer over later-developing embryos. These results can assist with patient counseling regarding expectations and outcomes. To our knowledge, this is the first study to examine embryonic euploidy as stratified by both day of blastocyst development and SART age group.8.
Sandrine Chamayou Maria Sicali Carmelita Alecci Carmen Ragolia Annalisa Liprino Daniela Nibali Giorgia Storaci Antonietta Cardea Antonino Guglielmino 《Journal of assisted reproduction and genetics》2017,34(4):479-486
Purpose
In a preimplantation genetic diagnosis for aneuploidy (PGD-A) program, the more embryos available for biopsy, consequently increases the chances of obtaining euploid embryos to transfer. The aim was to increase the number of viable euploid blastocysts in patients undergoing PGD-A using fresh oocytes together with previously accumulated vitrified oocytes.Methods
Sixty-nine patients with normal ovarian reserve underwent PGD-A for repeated implantation failure or recurrent pregnancy loss indication. After several cycles of ovarian stimulation, 591 accumulated vitrified oocytes and 463 fresh oocytes were micro-injected with the same partner’s semen sample. PGD-A was completed on 134 blastocysts from vitrified/warmed oocytes and 130 blastocysts from fresh oocytes.Results
A mean of 9.6% euploid blastocyst per micro-injected vitrified/warmed oocytes and 11.4% euploid blastocyst per micro-injected fresh oocyte were obtained (p?>?0.05). The euploidy and aneuploidy rates were comparable in blastocysts obtained from micro-injected vitrified/warmed oocytes and fresh oocytes (42.5 versus 40.8% and 57.5 versus 59.2%, p?>?0.05). Implantation rates of euploid blastocysts were comparable between the two sources of oocytes (56.0% from vitrified/warmed oocytes versus 60.9% from fresh oocytes, p?>?0.05).Conclusions
Oocyte vitrification and warming do not generate aneuploidy in blastocysts. The number of viable euploid embryos for transfer can be increased by using accumulated vitrified oocytes together with fresh oocytes in ICSI.Trial registration
NCT02820415 ClinicalTrials.gov9.
Jingye Zhang Wenrong Tao Hui Liu Guanling Yu Mei Li Shuiying Ma Keliang Wu 《Journal of assisted reproduction and genetics》2017,34(9):1173-1178
Purpose
This study aimed to test whether there is an association between embryo morphokinetic parameters and ploidy status.Methods
Patients with high risk of aneuploidy were analyzed by time-lapse microscopy combined with preimplantation genetic screening (PGS). Accordingly, 256 blastocysts from 75 patients were subjected to trophectoderm biopsy and microarray comparative genomic hybridization (array-CGH). Blastocyst development process was analyzed using time-lapse images.Results
Morphokinetic parameters: tPNf, t2, t3, t4, t5, t8, t9, tcom, tM, tSB, tB, tEB, CC1, CC2, CC3, S2, S3, t5-t2, and tB-tSB showed no significant difference in euploid embryos compared to aneuploid counterparts. In addition, two risk models based on previously published morphokinetic parameters failed to segregate euploid from aneuploid embryos.Conclusions
Morphokinetic parameters subjected to investigation in the present study failed to improve the chance of selecting euploid embryos.10.
John B. Whitney Mitchel C. Schiewe Robert E. Anderson 《Journal of assisted reproduction and genetics》2016,33(11):1507-1513
Purpose
The study aims to contrast the efficacy of trophectoderm biopsy preimplantation genetic screening (PGS)/vitrification (VTF)-all cycles to past treatment protocols. Specifically, do these applied technologies increase live birth rates on a per cycle/first transfer basis?Materials and methods
An observational, retrospective cohort study of first transfer outcomes was performed in two groups. Group 1 (PGS) included PGS/VTF-all cycles, and group 2 (no PGS) included the first transfer from non-PGS fresh cycles or VTF-ALL cycles. In group 1, all blastocysts were biopsied on days 5/6, vitrified and array CGH performed. Group 2 patients had embryo transfers on day 3 or day 5. All blastocysts were vitrified and warmed according to μS-VTF protocols. Clinical pregnancies and implantation were confirmed by ultrasound and live birth information attained. Results were stratified by age with donor cycles excluded, and to eliminate bias, the same groups were then validated on a per cycle basis. Chi-squared used to determine significance.Results
Analyzing 287 embryo transfers and 1,000+ PGS-tested blastocysts, an overall 97 % increase in live births favored group 1 (PGS). When utilizing PGS/VTF-ALL cycles, patients under 43 years old exhibited higher implantation, clinical pregnancy, and ongoing/live birth rates. Re-analyzing the data to include all cycles initiated revealed higher live birth rates in group 1 age groups ≤34 and 38–40 years old.Conclusion
Validating PGS on a per cycle basis eliminated data bias by including patients without blastocysts to biopsy or euploid embryos. Clearly, PGS uses blastocysts more efficiently to achieve success, while many women over 40 may benefit most by understanding why some failures occur.Support
None11.
Yi-zi Wang Chen-hui Ding Jing Wang Yan-hong Zeng Wen Zhou Rong Li Can-quan Zhou Ming-Fen Deng Yan-wen Xu 《Journal of assisted reproduction and genetics》2017,34(1):51-59
Purpose
The aim of this study is to investigate the minimum number of blastocysts for biopsy to increase the likelihood of obtaining at least one normal/balanced embryo in preimplantation genetic diagnosis (PGD) for translocation carriers.Methods
This blinded retrospective study included 55 PGD cycles for Robertsonian translocation (RT) and 181 cycles for reciprocal translocation (rcp) to indicate when only one of the couples carried a translocation. Single-nucleotide polymorphism microarray after trophectoderm biopsy was performed.Results
Reliable results were obtained for 355/379 (93.7 %) biopsied blastocysts in RT group and 986/1053 (93.6 %) in rcp group. Mean numbers of biopsied embryos per patient, normal/balanced embryos per patient, and mean normal/balanced embryo rate per patient were 7.4, 3.1, and 40.7 % in RT group and 8.0, 2.1, and 27.3 %, respectively, in rcp group. In a regression model, three factors significantly affected the number of genetically transferrable embryos: number of biopsied embryos (P?=?0.001), basal FSH level (P?=?0.040), and maternal age (P?=?0.027). ROC analysis with a cutoff of 1.5 was calculated for the number of biopsied embryos required to obtain at least one normal/balanced embryo for RT carriers. For rcp carriers, the cutoff was 3.5. The clinical pregnancy rate per embryo transfer was 44.2 and 42.6 % in RT and rcp groups (P?=?0.836).Conclusions
The minimum numbers of blastocysts to obtain at least one normal/balanced embryo for RT and rcp were 2 and 4 under the conditions of female age?<?37 years with a basal FSH level?<?11.4 IU/L.12.
Objective
To study the effect of GnRh agonist administration prior to estrogen–progesterone preparation of the endometrium on the implantation rate in frozen–thawed embryo transfer (FET) cycles in infertile patients treated with IVF/ICSI.Design
Prospective controlled study.Setting
Private center in Alexandria, Egypt.Patients
Patients undergoing frozen–thawed embryo transfer FET.Intervention(s)
Patients were divided into two groups, A and B. Group A patients consisted of 110 patients (110 cycles) who received daily subcutaneous injections of 0.1 mg of the GnRh agonist triptorelin starting from the mid-luteal phase of the cycle preceding the actual FET cycle. The dose was reduced to 0.05 mg from the second day of the cycle when daily oral estradiol valerate 6 mg was also started. Daily vaginal supplementation of micronized progesterone 400 mg b.d. was started after 12 days when the GnRh agonist was also stopped. Frozen–thawed embryos were transferred on day + 1 of their chronological age and when the endometrium reached 12 mm in thickness. Group B consisted of 100 patients (100 cycles) who started daily estradiol valerate 6 mg administration from the second day of the FET cycle and followed the same regimen but without prior treatment with triptorelin.Main Outcome Measures
Implantation and pregnancy rates were compared among the two groups.Results
There was a significant increase in implantation rate in the GnRh agonist group (group A) compared to the estrogen and progesterone only group (group B) (44.1 vs. 21.1 %; P = 0.002*). The pregnancy rate was also significantly higher in group A compared to group B (65.5 vs. 42 %, P = 0.013*).Conclusions
GnRh agonist administration during endometrial preparation for FET increases the implantation and pregnancy rates.13.
Liron Bar-El Yael Kalma Mira Malcov Tamar Schwartz Shaul Raviv Tania Cohen Hadar Amir Yoni Cohen Adi Reches Ami Amit Dalit Ben-Yosef 《Journal of assisted reproduction and genetics》2016,33(11):1449-1457
Purpose
The purpose of the study was to explore the effect of blastomere biopsy for preimplantation genetic diagnosis (PGD) on the embryos’ dynamics, further cleavage, development, and implantation.Methods
The study group included 366 embryos from all PGD treatments (September 2012 to June 2014) cultured in the EmbryoScope? time-lapse monitoring system. The control group included all intracytoplasmic sperm injection (ICSI) embryos cultured in EmbryoScope? until day 5 during the same time period (385 embryos). Time points of key embryonic events were analyzed with an EmbryoViewer?.Results
Most (88 %) of the embryos were biopsied at ≥8 cells. These results summarize the further dynamic development of the largest cohort of biopsied embryos and demonstrate that blastomere biopsy of cleavage-stage embryos significantly delayed compaction and blastulation compared to the control non-biopsied embryos. This delay in preimplanation developmental events also affected postimplantation development as observed when the dynamics of non-implanted embryos (known implantation data (KID) negative) were compared to those of implanted embryos (KID positive).Conclusion
Analysis of morphokinetic parameters enabled us to explore how blastomere biopsy interferes with the dynamic sequence of developmental events. Our results show that biopsy delays the compaction and the blastulation of the embryos, leading to a decrease in implantation.14.
Objoon Trachoo Chonthicha Satirapod Bhakbhoom Panthan Matchuporn Sukprasert Angkana Charoenyingwattana Wasun Chantratita Wicharn Choktanasiri Suradej Hongeng 《Journal of assisted reproduction and genetics》2017,34(1):109-116
Purpose
We aim to present a case of a healthy infant born after intracytoplasmic sperm injection-in vitro fertilization (ICSI-IVF) with a preimplantation genetic diagnosis (PGD) for pantothenate kinase-associated neurodegeneration (PKAN) due to PANK2 mutation.Methods
ICSI-IVF was performed on a Thai couple, 34-year-old female and 33-year-old male, with a family history of PKAN in their first child. Following fertilization, each of the embryos were biopsied in the cleavage stage and subsequently processed for whole-genome amplification. Genetic status of the embryos was diagnosed by linkage analysis and direct mutation testing using primer extension-based mini-sequencing. Comprehensive chromosomal aneuploidy screening was performed using a next-generation sequencing-based strategy.Results
Only a single cycle of ICSI-IVF was processed. There were seven embryos from this couple—two were likely affected, three were likely carriers, one was likely unaffected, and one failed in target genome amplification. Aneuploidy screening was performed before making a decision on embryo transfer, and only one unaffected embryo passed the screening. That embryo was transferred in a frozen thawed cycle, and the pregnancy was successful. The diagnosis was confirmed by amniocentesis, which presented with a result consistent with PGD. At 38 weeks of gestational age, a healthy male baby was born. Postnatal genetic confirmation was also consistent with PGD and the prenatal results. At the age of 24 months, the baby presented with normal growth and development lacking any neurological symptoms.Conclusions
We report the first successful trial of PGD for PKAN in a developing country using linkage analysis and mini-sequencing in cleavage stage embryos.15.
J. G. Lemmen N. M. Rodríguez L. D. Andreasen A. Loft S. Ziebe 《Journal of assisted reproduction and genetics》2016,33(7):849-854
Purpose
While stimulation of women prior to assisted reproduction is associated with increased success rates, the total biological pregnancy potential per stimulation cycle is rarely assessed.Methods
Retrospective sequential cohort study of the cumulative live birth rate in 1148 first IVF/ICSI-cycles and 5-year follow up of frozen embryo replacement (FER) cycles were used. Oocyte number, number of embryos transferred, and cryopreserved/thawed and transferred embryos in a FER cycle were registered for all patients. Children per oocyte and per transferred embryo and percentage of cycles with births were calculated.Results
We obtained 9529 oocytes. Embryos (2507) were transferred in either fresh or FER cycles, resulting in 422 births and 474 live born children. Median age of the women was 32.5 years (range 20–41.5 years). In total, 34.3 % of all cycles ended with a live birth while in 65.7 % of the cycles, no oocytes were capable of developing into a child. The average number of oocytes needed per live born child after transfer of fresh and thawed embryos was 20 as only 5.0 % of oocytes aspirated in the first IVF/ICSI cycle had the competence to develop into a child.Conclusions
In our setting, overall 5.0 % of the oocytes in a first cycle were biologically competent and in around 2/3 of all cycles, none of the oocytes had the potential to result in the birth of a child.16.
David Goodrich Tongji Xing Xin Tao Agnieszka Lonczak Yiping Zhan Jessica Landis Rebekah Zimmerman Richard T. ScottJr Nathan R. Treff 《Journal of assisted reproduction and genetics》2017,34(8):975-981
Purpose
A subset of preimplantation embryos identified as euploid may in fact possess both whole and sub-chromosomal mosaicism, raising concerns regarding the predictive value of current comprehensive chromosome screening (CCS) methods utilizing a single biopsy. Current CCS methods may be capable of detecting sub-chromosomal mosaicism in a trophectoderm biopsy by examining intermediate levels of segmental aneuploidy within a biopsy. This study evaluates the sensitivity and specificity of segmental aneuploidy detection by three commercially available CCS platforms utilizing a cell line mixture model of segmental mosaicism in a six-cell trophectoderm biopsy.Methods
Two cell lines with known karyotypes were obtained and mixed together at specific ratios of six total cells (0:6, 1:5, 2:4, 3:3, 4:2, 5:1, and 6:0). A female cell line containing a 16.2 Mb deletion on chromosome 5 and a male cell line containing a 25.5 Mb deletion on chromosome 4 were used to create mixtures at each level. Six replicates of each mixture were prepared, randomized, and blinded for analysis by one of the three CCS platforms (SNP-array, VeriSeq NGS, or NexCCS). Sensitivity and specificity of segmental aneuploidy at each level of mosaicism was determined and compared between each platform. Additionally, an alternative VeriSeq NGS analysis method utilizing previously published criteria was evaluated.Results
Examination of the default settings of each platform revealed that the sensitivity was significantly different between NexCCS and SNP up to 50% mosaicism, custom VeriSeq, and SNP-array up to 66% mosaicism, and between NexCCS and custom VeriSeq up to 50% mosaicism. However, no statistical difference was observed in mixtures with >50% mosaicism with any platform. No comparison was made between default VeriSeq, as it does not report segmental imbalances. Furthermore, while the use of previously published criteria for VeriSeq NGS significantly increased sensitivity at low levels of mosaicism, a significant decrease in specificity was observed (66% false positive prediction of segmental aneuploidy).Conclusion
These results demonstrate the potential of NGS-based detection methods to detect segmental mosaicism within a biopsy. However, these data also demonstrate that a balance between sensitivity and specificity should be more carefully considered. These results emphasize the importance of vigorous preclinical evaluation of new testing criteria prior to clinical implementation providing a point of departure for further algorithm development and improved detection of mosaicism within preimplantation embryos.17.
Jing Liu Xing Ling Wang Xiao Zhang Chun Yan Shen Zhan Zhang 《Journal of assisted reproduction and genetics》2016,33(3):373-378
Purpose
The aim of this study was to (1) investigate the incidence of embryos derived from “unfertilized oocytes” i.e., oocytes not displaying pronuclei (0PN) at the time of the fertilization check and (2) determine the clinical pregnancy rates when transferring 0PN-derived embryos.Methods
In this retrospective study, 4424 IVF-ET cycles were reviewed.Results
In total, 11.3 % (4966/43,949) 0PN-derived embryos were observed. It was found that female age, number of oocytes, and the top-quality embryo rate were significantly correlated with 0PN-derived embryo occurrence. The source of embryos transferred did not impact significantly on clinical pregnancy and live-birth rates. Of the 183 cycles included in this study where 275 0PN-derived embryos were transferred in total, only 0PN-derived embryos were available in 70 of those cycles. It was noteworthy that 13 healthy infants resulted from 0PN-derived embryos with an implantation rate of 17.0 %.Conclusion
These results indicate that the traditional method of excluding embryos because of those oocytes originally lacking any sign of a pronucleus at the fertilization check should be re-considered as transferring 0PN-derived embryos with subsequent expected developmental performance may be considered as an option for those patients where no other embryos are available.18.
Wen Li Yiyi Ma Shengqiang Yu Ningxia Sun Liang Wang Dongping Chen Guijiang Yang Sijia Lu Yangyang Li Bo Yang Changlin Mei 《Journal of assisted reproduction and genetics》2017,34(12):1653-1658
Background
Autosomal dominant polycystic kidney disease (ADPKD, autosomal dominant PKD or adult-onset PKD) is the most prevalent and potentially lethal kidney disease that is hereditary and lacks effective treatment. Preimplantation genetic diagnosis (PGD) of embryos in assistant reproductive technology (ART) helps to select mutation-free embryos for blocking ADPKD inheritance from the parents to their offspring. However, there are multiple pseudogenes in the PKD1 coding region, which make blocking ADPKD inheritance by PGD complicated and difficult. Therefore, this technique has not been recommended and used routinely to ADPKD family plan.Methods and Results
Here, we report a new strategy of performing PGD in screening (target-) mutation-free embryos. We firstly used a long-range PCR amplification and next generation sequencing to identify the potential PKD1 mutant(s). After pathogenic variants were detected, multiple annealing and looping-based amplification cycles (MALBAC), a recently developed whole genome amplification method, was used to screen embryo cells. We successfully distinguished the mutated allele among pseudogenes and obtained mutation-free embryos for implantation. The first embryo transfer attempt resulted in a healthy live birth free of ADPKD condition and chromosomal anomalies which was confirmed by aminocentesis at week 18 of gestation, and by performing live birth genetic screening.Conclusions
The first MALBAC-PGD attempt in ADPKD patient resulted in a healthy live birth free of ADPKD and chromosomal anomalies. MALBAC-PGD also enables selecting embryos without aneuploidy together and target gene mutation, thereby increasing implantation and live birth rates.19.
Katarzyna Litwicka Cecilia Mencacci Cristiana Arrivi Maria Teresa Varricchio Alina Caragia Maria Giulia Minasi Ermanno Greco 《Journal of assisted reproduction and genetics》2018,35(3):449-455
Purpose
The aim of the present study was to evaluate whether in a modified natural cycle (modified-NC) for a frozen-thawed single euploid blastocyst transfer, a critical LH value, above which human chorionic gonadotropin (hCG) administration should be avoided, may be defined.Methods
One hundred and sixty-seven patients underwent modified natural cycle in order to transfer a single frozen-thawed euploid blastocyst. All embryos were obtained by intracytoplasmic sperm injection and were biopsied at the blastocyst stage and analyzed by means of array comparative genomic hybridization (aCGH). Ovulation was induced using 10.000 IU hCG when the mean follicle diameter was at least of 17 mm, independently from LH values. The primary end points were the hCG-positive test and clinical pregnancy. The interim analysis showed that LH value ≥?13 mIU/ml on the day of hCG injection may negatively influence the clinical results, suggesting that in this condition, it should be advisable waiting for spontaneous ovulation.Results
Among patients who received hCG for ovulation induction, the hCG-positive test and clinical pregnancy rates in modified-NC were significantly lower in cycles with LH?≥?13 mIU/ml in respect to those with LH?<?13 mIU/ml (45.4 vs 73.3 and 36.4 vs 65.9%, in LH?≥?13 and LH?<?13 groups, respectively). In patients with LH value ≥?13 mIU/ml, hCG administration led to significantly lower rates of hCG-positive test (45.4 vs 74.5% in hCG administration and spontaneous ovulation groups, respectively) and clinical pregnancy (36.4 vs 64.7% in hCG administration and spontaneous ovulation groups, respectively). The baseline patient characteristics were comparable in all groups.Conclusions
The findings of this study highlight that LH elevation ≥?13 mIU/ml prior to hCG administration may negatively affect clinical pregnancy rates in modified-NC for single euploid blastocyst transfer. The LH determination should be routinely performed during follicular monitoring. In the presence of LH level ≥?13 mIU/ml, hCG administration should be avoided, and the embryo transfer should be planned only after spontaneous follicular rupture.20.
Kemal Ozgur Murat Berkkanoglu Hasan Bulut Peter Humaidan Kevin Coetzee 《Journal of assisted reproduction and genetics》2016,33(2):207-214