{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,6,12]],"date-time":"2026-06-12T01:29:15Z","timestamp":1781227755334,"version":"3.54.1"},"reference-count":44,"publisher":"MDPI AG","issue":"18","license":[{"start":{"date-parts":[[2022,9,16]],"date-time":"2022-09-16T00:00:00Z","timestamp":1663286400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>Preimplantation genetic testing (PGT) is widely used to select unaffected embryos, increasing the odds of having a healthy baby. During the last few decades, it was accepted that monozygotic dichorionic diamniotic twin pregnancies occurred from the embryo splitting before Day 3 postfertilization according to Corner\u2019s dogma. Hence, the occurrence of a dichorionic diamniotic twin pregnancy after a single blastocyst transfer was considered a dizygotic pregnancy resulting from blastocyst transfer and concurrent natural fertilization. In our study, we have provided for the first time molecular proof that a single blastocyst transfer can result in a monozygotic dichorionic diamniotic twin pregnancy, invalidating Corner\u2019s dogma. In this case, we recommend systematically assessing the genetic status of dichorionic twins after single blastocyst transfer using prenatal diagnosis to exclude the risk from a potential concurrent spontaneous pregnancy and to ensure that both fetuses are unaffected. To achieve this goal, we have developed here an innovative noninvasive prenatal diagnosis by exclusion of paternal variants with droplet digital PCR, maximizing the reliability of genetic diagnosis. Further multicentric prospective studies using genetic testing are now required to establish the rate of blastocyst splitting leading to dichorionic pregnancy in PGT and to identify the risk factors.<\/jats:p>","DOI":"10.3390\/ijms231810835","type":"journal-article","created":{"date-parts":[[2022,9,18]],"date-time":"2022-09-18T23:39:22Z","timestamp":1663544362000},"page":"10835","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":5,"title":["Molecular Characterization of a Rare Case of Monozygotic Dichorionic Diamniotic Twin Pregnancy after Single Blastocyst Transfer in Preimplantation Genetic Testing (PGT)"],"prefix":"10.3390","volume":"23","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-3354-1551","authenticated-orcid":false,"given":"Sophie","family":"Brouillet","sequence":"first","affiliation":[{"name":"DEFE, University of Montpellier, INSERM, Montpellier, France"},{"name":"Biologie de la Reproduction\/DPI et CECOS, D\u00e9partement de Biologie de la Reproduction, CHU of Montpellier, Montpellier, France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sandie","family":"Mereuze","sequence":"additional","affiliation":[{"name":"Laboratory of Molecular Genetics, CHU of Montpellier, Montpellier, France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1956-8571","authenticated-orcid":false,"given":"No\u00e9mie","family":"Ranisavljevic","sequence":"additional","affiliation":[{"name":"Department of Reproductive Medicine, CHU of Montpellier, Montpellier, France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Claire","family":"Chauveau","sequence":"additional","affiliation":[{"name":"Department of Medical Genetics, CHU of Montpellier, Montpellier, France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Samir","family":"Hamamah","sequence":"additional","affiliation":[{"name":"DEFE, University of Montpellier, INSERM, Montpellier, France"},{"name":"Biologie de la Reproduction\/DPI et CECOS, D\u00e9partement de Biologie de la Reproduction, CHU of Montpellier, Montpellier, France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Julie","family":"Cattin","sequence":"additional","affiliation":[{"name":"Service de Gyn\u00e9cologie-Obst\u00e9trique, CHU Jean Minjoz, Besan\u00e7on, France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0524-4827","authenticated-orcid":false,"given":"Camille","family":"Verebi","sequence":"additional","affiliation":[{"name":"Service de M\u00e9decine G\u00e9nomique, Maladies de Syst\u00e8me et d\u2019Organe, F\u00e9d\u00e9ration de G\u00e9n\u00e9tique et de M\u00e9decine G\u00e9nomique, DMU BioPhyGen, APHP Centre\u2014Universit\u00e9 Paris Cit\u00e9, H\u00f4pital Cochin, Paris, France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Christelle","family":"Cabrol","sequence":"additional","affiliation":[{"name":"Centre de G\u00e9n\u00e9tique Humaine, CHU Jean-Minjoz, Besan\u00e7on, France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Aliya","family":"Ishmukhametova","sequence":"additional","affiliation":[{"name":"Laboratory of Molecular Genetics, CHU of Montpellier, Montpellier, France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Anne","family":"Girardet","sequence":"additional","affiliation":[{"name":"Laboratory of Molecular Genetics, CHU of Montpellier, Montpellier, France"},{"name":"PhyMedExp, CHU of Montpellier, University of Montpellier, INSERM, CNRS, Montpellier, France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Tal","family":"Anahory","sequence":"additional","affiliation":[{"name":"Department of Reproductive Medicine, CHU of Montpellier, Montpellier, France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2959-0935","authenticated-orcid":false,"given":"Marjolaine","family":"Willems","sequence":"additional","affiliation":[{"name":"Reference Centre AD SOOR, AnDDI-RARE, INSERM U1298, INM, Department of Medical Genetics, Arnaud de Villeneuve Hospital and University of Montpellier, Montpellier, France"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2022,9,16]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"160","DOI":"10.1163\/15718093-12420347","article-title":"Preimplantation Genetic Diagnosis: The Situation in France and in Other European Countries","volume":"24","author":"Duguet","year":"2017","journal-title":"Eur. J. Health Law"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"a036681","DOI":"10.1101\/cshperspect.a036681","article-title":"Regulating Preimplantation Genetic Testing across the World: A Comparison of International Policy and Ethical Perspectives","volume":"10","author":"Ginoza","year":"2019","journal-title":"Cold Spring Harb. Perspect. Med."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"41","DOI":"10.1016\/j.rbms.2017.01.001","article-title":"Comparative preimplantation genetic diagnosis policy in Europe and the USA and its implications for reproductive tourism","volume":"3","author":"Bayefsky","year":"2016","journal-title":"Reprod. Biomed. Soc. Online"},{"key":"ref_4","unstructured":"The European IVF-Monitoring Consortium (EIM) for the European Society of Human Reproduction and Embryology (ESHRE), Wyns, C., De Geyter, C., Calhaz-Jorge, M., Kupka, M.S., Motrenko, T., Smeenk, J., Bergh, C., Tandler-Schneider, A., and Rugescu, I.A. (2021). ART in Europe, 2017: Results generated from European registries by ESHRE. Hum. Reprod. Open, 2021, hoab026."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"468","DOI":"10.1093\/humupd\/dmy006","article-title":"Determinants of monozygotic twinning in ART: A systematic review and a meta-analysis","volume":"24","author":"Hviid","year":"2018","journal-title":"Hum. Reprod. Updat."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"178","DOI":"10.1016\/S0015-0282(00)00557-4","article-title":"Do assisted reproductive technologies have effects on the demography of monozygotic twinning?","volume":"74","author":"Saito","year":"2000","journal-title":"Fertil. Steril."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"72","DOI":"10.1017\/thg.2015.96","article-title":"Risk of Monozygotic Twins After Assisted Reproduction: A Population-Based Approach","volume":"19","author":"Parazzini","year":"2016","journal-title":"Twin Res. Hum. Genet."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"302","DOI":"10.1016\/j.fertnstert.2018.10.025","article-title":"Risk factors for monozygotic twinning after in vitro fertilization: A systematic review and meta-analysis","volume":"111","author":"Busnelli","year":"2019","journal-title":"Fertil. Steril."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"1984","DOI":"10.1093\/humrep\/dey294","article-title":"Prevalence and risk factors of zygotic splitting after 937 848 single embryo transfer cycles","volume":"33","author":"Ikemoto","year":"2018","journal-title":"Hum. Reprod."},{"key":"ref_10","doi-asserted-by":"crossref","unstructured":"Li, H., Shen, T., and Sun, X. (2020). Monozygotic dichorionic-diamniotic pregnancies following single frozen-thawed blastocyst transfer: A retrospective case series. BMC Pregnancy Childbirth, 20.","DOI":"10.1186\/s12884-020-03450-5"},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"933","DOI":"10.1016\/0002-9378(55)90001-6","article-title":"The observed embryology of human single-ovum twins and other multiple births","volume":"70","author":"Corner","year":"1955","journal-title":"Am. J. Obstet. Gynecol."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"502","DOI":"10.1016\/j.fertnstert.2009.03.064","article-title":"Monozygotic twinning: An eight-year experience at a large IVF center","volume":"94","author":"Knopman","year":"2010","journal-title":"Fertil. Steril."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"827","DOI":"10.1017\/thg.2013.32","article-title":"The Precise Timing of Embryo Splitting for Monozygotic Dichorionic Diamniotic Twins: When Does Embryo Splitting for Monozygotic Dichorionic Diamniotic Twins Occur? Evidence for Splitting at the Morula\/Blastocyst Stage from Studies of In Vitro Fertilization","volume":"16","author":"Kyono","year":"2013","journal-title":"Twin Res. Hum. Genet."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"2109","DOI":"10.1007\/s10815-018-1329-8","article-title":"Multi-chorionic pregnancies following single embryo transfer at the blastocyst stage: A case series and review of the literature","volume":"35","author":"Sundaram","year":"2018","journal-title":"J. Assist. Reprod. Genet."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"735","DOI":"10.1016\/S0140-6736(03)14237-7","article-title":"Twinning","volume":"362","author":"Hall","year":"2003","journal-title":"Lancet"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"1185","DOI":"10.1016\/j.fertnstert.2015.01.033","article-title":"Monozygotic twinning after assisted reproductive technologies: A case report of asymmetric development and incidence during 19 years in an international group of in vitro fertilization clinics","volume":"103","author":"Tocino","year":"2015","journal-title":"Fertil. Steril."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"51","DOI":"10.1017\/thg.2020.5","article-title":"The Incidence of Dichorionic Diamniotic Twin Pregnancy after Single Blastocyst Embryo Transfer and Zygosity: 8 Years of Single-Center Experience","volume":"23","author":"Konno","year":"2020","journal-title":"Twin Res. Hum. Genet."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"1438","DOI":"10.1093\/humrep\/deu064","article-title":"Incidence and zygosity of twin births following transfers using a single fresh or frozen embryo","volume":"29","author":"Osianlis","year":"2014","journal-title":"Hum. Reprod."},{"key":"ref_19","doi-asserted-by":"crossref","unstructured":"Knopman, J.M., Krey, L.C., Oh, C., Lee, J., McCaffrey, C., and Noyes, N. (2013). Withdrawn: What makes them split? Identifying risk factors that lead to monozygotic twins after in vitro fertilization. Fertil. Steril.","DOI":"10.1016\/j.fertnstert.2013.08.053"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"1155","DOI":"10.1016\/S0015-0282(00)00518-5","article-title":"Blastocyst score affects implantation and pregnancy outcome: Towards a single blastocyst transfer","volume":"73","author":"Gardner","year":"2000","journal-title":"Fertil. Steril."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"182","DOI":"10.1111\/j.1471-0528.2002.01172.x","article-title":"Prediction of chorionicity in twin pregnancies at 10\u201314 weeks of gestation","volume":"109","author":"Soothill","year":"2002","journal-title":"BJOG"},{"key":"ref_22","doi-asserted-by":"crossref","unstructured":"Wang, C.-Y., Tang, Y.-A., Lee, I.-W., Chang, F.-M., Chien, C.-W., Pan, H.-A., and Sun, H.S. (2021). Development and validation of an expanded targeted sequencing panel for non-invasive prenatal diagnosis of sporadic skeletal dysplasia. BMC Med Genom., 14.","DOI":"10.1186\/s12920-021-01063-1"},{"key":"ref_23","doi-asserted-by":"crossref","unstructured":"Pacault, M., Verebi, C., Lopez, M., Vaucouleur, N., Orhant, L., Deburgrave, N., Leturcq, F., Vidaud, D., Girodon, E., and Bienvenu, T. (2022). Non-invasive prenatal diagnosis of single gene disorders by paternal mutation exclusion: 3 years of clinical experience. BJOG Int. J. Obstet. Gynaecol.","DOI":"10.1111\/1471-0528.17201"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"409","DOI":"10.1002\/pd.2897","article-title":"Amniocentesis in twin pregnancies: A systematic review of the literature","volume":"32","author":"Vink","year":"2011","journal-title":"Prenat. Diagn."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"647","DOI":"10.1002\/uog.22143","article-title":"Risk of fetal loss following amniocentesis or chorionic villus sampling in twin pregnancy: Systematic review and meta-analysis","volume":"56","author":"Khalil","year":"2020","journal-title":"Ultrasound Obstet. Gynecol."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"833","DOI":"10.1016\/S0015-0282(16)47894-5","article-title":"Monozygotic twins from in vitro fertilization","volume":"41","author":"Yovich","year":"1984","journal-title":"Fertil. Steril."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"284","DOI":"10.1016\/j.fertnstert.2017.11.003","article-title":"Successful quintuplet pregnancy of monochorionic male quadruplets and single female after double embryo transfer: Case report and review of the literature","volume":"109","author":"Schlueter","year":"2018","journal-title":"Fertil. Steril."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"315","DOI":"10.1530\/jrf.0.0690315","article-title":"Effect of various procedures on the viability of mouse embryos containing half the normal number of blastomeres","volume":"69","author":"Tsunoda","year":"1983","journal-title":"Reproduction"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"298","DOI":"10.1038\/277298a0","article-title":"A method for culture of micromanipulated sheep embryos and its use to produce monozygotic twins","volume":"277","author":"Willadsen","year":"1979","journal-title":"Nat."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"23","DOI":"10.1016\/S0093-691X(81)80015-5","article-title":"The production of monozygotic twins of preselected parentage by micromanipulation of non-surgically collected cow embryos","volume":"15","author":"Willadsen","year":"1981","journal-title":"Theriogenology"},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"1742","DOI":"10.1093\/humrep\/den190","article-title":"The four blastomeres of a 4-cell stage human embryo are able to develop individually into blastocysts with inner cell mass and trophectoderm","volume":"23","author":"Cauffman","year":"2008","journal-title":"Hum. Reprod."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"27","DOI":"10.1017\/S0967199413000257","article-title":"The timing of monozygotic twinning: A criticism of the common model","volume":"23","author":"Herranz","year":"2013","journal-title":"Zygote"},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"312","DOI":"10.1017\/S0967199413000579","article-title":"Comment on G. Herranz: The timing of monozygotic twinning: A criticism of the common model","volume":"23","author":"Denker","year":"2015","journal-title":"Zygote"},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"255","DOI":"10.1007\/s10815-011-9707-5","article-title":"A successful birth of healthy monozygotic dichorionic diamniotic (DD) twins of the same gender following a single vitrified-warmed blastocyst transfer","volume":"29","author":"Shibuya","year":"2012","journal-title":"J. Assist. Reprod. Genet."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"1047","DOI":"10.1016\/S0015-0282(00)01554-5","article-title":"Atypical hatching of a human blastocyst leading to monozygotic twinning: A case report","volume":"74","author":"Wyns","year":"2000","journal-title":"Fertil. Steril."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"118","DOI":"10.1016\/j.fertnstert.2017.10.003","article-title":"Not all twins are monozygotic after elective single embryo transfer: Analysis of 32,600 elective single embryo transfer cycles as reported to the Society for Assisted Reproductive Technology","volume":"109","author":"Vega","year":"2018","journal-title":"Fertil. Steril."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"656","DOI":"10.1002\/uog.22041","article-title":"New approach for estimating risk of miscarriage after chorionic villus sampling","volume":"56","author":"Gil","year":"2020","journal-title":"Ultrasound Obstet. Gynecol."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"442","DOI":"10.1002\/uog.20353","article-title":"Risk of miscarriage following amniocentesis or chorionic villus sampling: Systematic review of literature and updated meta-analysis","volume":"54","author":"Salomon","year":"2019","journal-title":"Ultrasound Obstet. Gynecol."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"383","DOI":"10.1111\/cge.13680","article-title":"Integrated analysis of COL2A1 variant data and classification of type II collagenopathies","volume":"97","author":"Zhang","year":"2019","journal-title":"Clin. Genet."},{"key":"ref_40","first-page":"992","article-title":"The expanding spectrum of COL2A1 gene variants IN 136 patients with a skeletal dysplasia phenotype","volume":"24","author":"Dumont","year":"2015","journal-title":"Eur. J. Hum. Genet."},{"key":"ref_41","doi-asserted-by":"crossref","unstructured":"ESHRE PGT-M Working Group, Carvalho, F., Moutou, C., Dimitriadou, E., Dreesen, J., Gim\u00e9nez, C., Goossens, V., Kakourou, G., Vermeulen, N., and Zuccarello, D. (2020). ESHRE PGT Consortium good practice recommendations for the detection of monogenic disorders. Hum. Reprod. Open, 2020, hoaa018.","DOI":"10.1093\/hropen\/hoaa018"},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"399","DOI":"10.1093\/molehr\/gag046","article-title":"The minisequencing method: An alternative strategy for preimplantation genetic diagnosis of single gene disorders","volume":"9","author":"Fiorentino","year":"2003","journal-title":"Mol. Hum. Reprod."},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"9389","DOI":"10.1073\/pnas.86.23.9389","article-title":"Single-sperm typing: Determination of genetic distance between the G gamma-globin and parathyroid hormone loci by using the polymerase chain reaction and allele-specific oligomers","volume":"86","author":"Cui","year":"1989","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"728","DOI":"10.1515\/cclm-2017-0689","article-title":"Non-invasive prenatal diagnosis of paternally inherited disorders from maternal plasma: Detection of NF1 and CFTR mutations using droplet digital PCR","volume":"56","author":"Gruber","year":"2018","journal-title":"Clin. Chem. Lab. Med."}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/23\/18\/10835\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T00:33:05Z","timestamp":1760142785000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/23\/18\/10835"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2022,9,16]]},"references-count":44,"journal-issue":{"issue":"18","published-online":{"date-parts":[[2022,9]]}},"alternative-id":["ijms231810835"],"URL":"https:\/\/doi.org\/10.3390\/ijms231810835","relation":{},"ISSN":["1422-0067"],"issn-type":[{"value":"1422-0067","type":"electronic"}],"subject":[],"published":{"date-parts":[[2022,9,16]]}}}