Danilo Cimadomo1,2,3,4, Laura Rienzi1,2,3,4, Valeria Romanelli5, Erminia Alviggi2, Paolo Emanuele Levi-Setti6, Elena Albani6, Ludovica Dusi3, Letizia Papini4, Claudia Livi7, Francesca Benini7, Antonella Smeraldi6, Cristina Patassini5, Filippo Maria Ubaldi1,2,3,4, Antonio Capalbo5,8. 1. Clinica Valle Giulia, G.en.e.r.a. Centers for Reproductive Medicine, Via Giuseppe de Notaris 2b, Rome, Italy. 2. Clinica Ruesch, G.en.e.r.a. Centers for Reproductive Medicine, Viale Maria Cristina di Savoia 39, Naples, Italy. 3. G.en.e.r.a. Veneto, G.en.e.r.a. Centers for Reproductive Medicine, Via Fermi 1, Marostica, Italy. 4. G.en.e.r.a. Umbria, G.en.e.r.a. Centers for Reproductive Medicine, Via Papa Paolo VI, Umbertide, Italy. 5. Igenomix, Via Fermi 1, 36063 Marostica, Italy. 6. Humanitas Research Hospital, Humanitas Fertility Center, Via Alessandro Manzoni 56, Rozzano, Italy. 7. Centro di Procreazione Medicalmente Assistita Demetra, Via Giulio Caccini 18, Florence, Italy. 8. Dipartimento di Scienze Anatomiche, Istologiche, Medico-legali e Dell'apparato Locomotore, Sezione di Istologia ed Embriologia Medica, University 'Sapienza' of Rome, Via Scarpa 16, Rome, Italy.
Abstract
STUDY QUESTION: Can a second round of biopsy, vitrification and chromosomal testing provide a valid diagnosis where the first attempt fails? SUMMARY ANSWER: The risk of inconclusive chromosomal-assessment after trophectoderm biopsy was 2.5% but a further biopsy and vitrification-warming appeared not to impair the competence of euploid blastocysts. WHAT IS KNOWN ALREADY: The increasing implementation of multicell trophectoderm biopsy has significantly reduced the risk of inconclusive diagnosis after preimplantation-genetic-testing (PGT). Yet, few reports have defined the variables that influence the risk of failure or described the technical and clinical outcomes after re-biopsy. STUDY DESIGN, SIZE, DURATION: Retrospective multicenter study involving 8990 blastocyst biopsies conducted between April 2013 and September 2017 at six IVF centers but analyzed at a single genetic laboratory. A total of 206 blastocysts were successfully re-biopsied after warming and re-expansion, then re-vitrified. And 49 of these blastocysts were diagnosed euploid and used in single-embryo-transfers (SETs). Logistic regression analyses were conducted. PARTICIPANTS/MATERIALS, SETTING, METHODS: A total of 3244 PGT-for-aneuploidies (PGT-A) cycles with a freeze-all approach, vitrification and qPCR-based analysis were performed by 2687 consenting couples. DNA amplification failure (AF) or non-concurrent data resulted in inconclusive diagnoses. In case of DNA amplification, the cellularity of the biopsy was estimated according to a previously validated method. Euploid SETs were performed. Clinical pregnancy, miscarriage, live birth rates (LBR) and perinatal outcomes were monitored. MAIN RESULTS AND THE ROLE OF CHANCE: Overall, 2.5% of trophectoderm biopsies resulted in an inconclusive diagnosis (N = 228/8990). Specifically, 2% (N = 176/8990) resulted in AF and 0.5% (N = 52/8990) in non-concurrent results. The only parameters significantly associated with inconclusive diagnoses were the IVF center and the embryo age (days) at biopsy. Among samples with successful amplification, the number of cells in the biopsy and the day of biopsy were critical to limit non-concurrent results. In total, 213 blastocysts with an inconclusive diagnosis were warmed for re-analysis and the survival rate was 96.7% (N = 206/213). The euploidy rate in blastocysts biopsied twice was 51.9% (N = 107/206) and the euploid embryos were re-vitrified. Overall, 49 euploid embryos were warmed for replacement and all survived. The LBR after SET was 38.8% (N = 19/49). No minor/major obstetrical/perinatal complication was reported. LIMITATIONS, REASONS FOR CAUTION: A single aneuploidy-testing method was adopted in this retrospective analysis. A more powered report of the clinical and obstetrical/perinatal outcomes after re-biopsied and re-vitrified blastocysts euploid SET requires a larger sample size. WIDER IMPLICATIONS OF THE FINDINGS: It is important to re-biopsy and re-vitrify undiagnosed blastocysts since healthy live births can result from them. STUDY FUNDING/COMPETING INTEREST(S): None. TRIAL REGISTRATION NUMBER: None.
STUDY QUESTION: Can a second round of biopsy, vitrification and chromosomal testing provide a valid diagnosis where the first attempt fails? SUMMARY ANSWER: The risk of inconclusive chromosomal-assessment after trophectoderm biopsy was 2.5% but a further biopsy and vitrification-warming appeared not to impair the competence of euploid blastocysts. WHAT IS KNOWN ALREADY: The increasing implementation of multicell trophectoderm biopsy has significantly reduced the risk of inconclusive diagnosis after preimplantation-genetic-testing (PGT). Yet, few reports have defined the variables that influence the risk of failure or described the technical and clinical outcomes after re-biopsy. STUDY DESIGN, SIZE, DURATION: Retrospective multicenter study involving 8990 blastocyst biopsies conducted between April 2013 and September 2017 at six IVF centers but analyzed at a single genetic laboratory. A total of 206 blastocysts were successfully re-biopsied after warming and re-expansion, then re-vitrified. And 49 of these blastocysts were diagnosed euploid and used in single-embryo-transfers (SETs). Logistic regression analyses were conducted. PARTICIPANTS/MATERIALS, SETTING, METHODS: A total of 3244 PGT-for-aneuploidies (PGT-A) cycles with a freeze-all approach, vitrification and qPCR-based analysis were performed by 2687 consenting couples. DNA amplification failure (AF) or non-concurrent data resulted in inconclusive diagnoses. In case of DNA amplification, the cellularity of the biopsy was estimated according to a previously validated method. Euploid SETs were performed. Clinical pregnancy, miscarriage, live birth rates (LBR) and perinatal outcomes were monitored. MAIN RESULTS AND THE ROLE OF CHANCE: Overall, 2.5% of trophectoderm biopsies resulted in an inconclusive diagnosis (N = 228/8990). Specifically, 2% (N = 176/8990) resulted in AF and 0.5% (N = 52/8990) in non-concurrent results. The only parameters significantly associated with inconclusive diagnoses were the IVF center and the embryo age (days) at biopsy. Among samples with successful amplification, the number of cells in the biopsy and the day of biopsy were critical to limit non-concurrent results. In total, 213 blastocysts with an inconclusive diagnosis were warmed for re-analysis and the survival rate was 96.7% (N = 206/213). The euploidy rate in blastocysts biopsied twice was 51.9% (N = 107/206) and the euploid embryos were re-vitrified. Overall, 49 euploid embryos were warmed for replacement and all survived. The LBR after SET was 38.8% (N = 19/49). No minor/major obstetrical/perinatal complication was reported. LIMITATIONS, REASONS FOR CAUTION: A single aneuploidy-testing method was adopted in this retrospective analysis. A more powered report of the clinical and obstetrical/perinatal outcomes after re-biopsied and re-vitrified blastocysts euploid SET requires a larger sample size. WIDER IMPLICATIONS OF THE FINDINGS: It is important to re-biopsy and re-vitrify undiagnosed blastocysts since healthy live births can result from them. STUDY FUNDING/COMPETING INTEREST(S): None. TRIAL REGISTRATION NUMBER: None.
Authors: Laura Girardi; Munevver Serdarogullari; Cristina Patassini; Maurizio Poli; Marco Fabiani; Silvia Caroselli; Onder Coban; Necati Findikli; Fazilet Kubra Boynukalin; Mustafa Bahceci; Rupali Chopra; Rita Canipari; Danilo Cimadomo; Laura Rienzi; Filippo Ubaldi; Eva Hoffmann; Carmen Rubio; Carlos Simon; Antonio Capalbo Journal: Am J Hum Genet Date: 2020-03-26 Impact factor: 11.025
Authors: Sandro C Esteves; Hakan Yarali; Filippo M Ubaldi; José F Carvalho; Fabiola C Bento; Alberto Vaiarelli; Danilo Cimadomo; İrem Y Özbek; Mehtap Polat; Gurkan Bozdag; Laura Rienzi; Carlo Alviggi Journal: Front Endocrinol (Lausanne) Date: 2020-01-24 Impact factor: 5.555