{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,10,29]],"date-time":"2025-10-29T03:52:04Z","timestamp":1761709924668,"version":"build-2065373602"},"reference-count":31,"publisher":"MDPI AG","issue":"12","license":[{"start":{"date-parts":[[2021,11,25]],"date-time":"2021-11-25T00:00:00Z","timestamp":1637798400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Genes"],"abstract":"<jats:p>Mutations in the HSD17B3 gene cause HSD17B3 deficiency and result in 46, XY Disorders of Sex Development (46, XY DSD). The diagnosis of 46, XY DSD is very challenging and not rarely is confirmed only at older ages, when an affected XY female presents with primary amenorrhea or develops progressive virilization. The patient described in this paper represents a case of discrepancies between non-invasive prenatal testing (NIPT) and ultrasound based fetal sex determination detected during prenatal screening. Exome sequencing was performed on the cell free fetal DNA (cffDNA), amniotic fluid, and the parents. Libraries were generated according to the manufacturer\u2019s protocols using TruSight One Kits (Illumina Inc., San Diego, CA, USA). Sequencing was carried out on NEXT Seq 500 (Illumina) to mean sequencing depth of at least 100\u00d7. A panel of sexual disease genes was used in order to search for a causative variant. The finding of a mutation (c.645 A&gt;T, p.Glu215Asp) in HSD17B3 gene in amniotic fluid as well as in cffDNA and both parents supported the hypothesis of the HSD17B3 deficiency. In conclusion, we used clinical exome sequencing and non-invasive prenatal detection, providing a solution for NIPT of a single-gene disorder. Early genetic diagnoses are useful for patients and clinicians, contribute to clinical knowledge of DSD, and are invaluable for genetic counseling of couples contemplating future pregnancies.<\/jats:p>","DOI":"10.3390\/genes12121890","type":"journal-article","created":{"date-parts":[[2021,11,29]],"date-time":"2021-11-29T05:23:02Z","timestamp":1638163382000},"page":"1890","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":7,"title":["Detection of 46, XY Disorder of Sex Development (DSD) Based on Plasma Cell-Free DNA and Targeted Next-Generation Sequencing"],"prefix":"10.3390","volume":"12","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-0553-2087","authenticated-orcid":false,"given":"Luigia","family":"De Falco","sequence":"first","affiliation":[{"name":"AMES, Centro Polidiagnostico Strumentale, 80013 Naples, Italy"},{"name":"Fondazione Genetica per la Vita Onlus, Via Cuma, 80132 Naples, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Carmelo","family":"Piscopo","sequence":"additional","affiliation":[{"name":"Medical and Laboratory Genetic Unit, A. Cardarelli Hospital, 80131 Naples, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Rossana","family":"D\u2019Angelo","sequence":"additional","affiliation":[{"name":"AMES, Centro Polidiagnostico Strumentale, 80013 Naples, Italy"},{"name":"Fondazione Genetica per la Vita Onlus, Via Cuma, 80132 Naples, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Eloisa","family":"Evangelista","sequence":"additional","affiliation":[{"name":"AMES, Centro Polidiagnostico Strumentale, 80013 Naples, Italy"},{"name":"Fondazione Genetica per la Vita Onlus, Via Cuma, 80132 Naples, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Teresa","family":"Suero","sequence":"additional","affiliation":[{"name":"AMES, Centro Polidiagnostico Strumentale, 80013 Naples, Italy"},{"name":"Fondazione Genetica per la Vita Onlus, Via Cuma, 80132 Naples, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Roberto","family":"Sirica","sequence":"additional","affiliation":[{"name":"AMES, Centro Polidiagnostico Strumentale, 80013 Naples, Italy"},{"name":"Fondazione Genetica per la Vita Onlus, Via Cuma, 80132 Naples, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Raffaella","family":"Ruggiero","sequence":"additional","affiliation":[{"name":"AMES, Centro Polidiagnostico Strumentale, 80013 Naples, Italy"},{"name":"Fondazione Genetica per la Vita Onlus, Via Cuma, 80132 Naples, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Giovanni","family":"Savarese","sequence":"additional","affiliation":[{"name":"AMES, Centro Polidiagnostico Strumentale, 80013 Naples, Italy"},{"name":"Fondazione Genetica per la Vita Onlus, Via Cuma, 80132 Naples, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Antonella","family":"Di Carlo","sequence":"additional","affiliation":[{"name":"AMES, Centro Polidiagnostico Strumentale, 80013 Naples, Italy"},{"name":"Fondazione Genetica per la Vita Onlus, Via Cuma, 80132 Naples, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Giulia","family":"Furino","sequence":"additional","affiliation":[{"name":"AMES, Centro Polidiagnostico Strumentale, 80013 Naples, Italy"},{"name":"Fondazione Genetica per la Vita Onlus, Via Cuma, 80132 Naples, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Ciro","family":"Scarpato","sequence":"additional","affiliation":[{"name":"Ambulatorio Medicina Prenatale, PO S. Giuliano, 80014 Naples, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Antonio","family":"Fico","sequence":"additional","affiliation":[{"name":"AMES, Centro Polidiagnostico Strumentale, 80013 Naples, Italy"},{"name":"Fondazione Genetica per la Vita Onlus, Via Cuma, 80132 Naples, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2021,11,25]]},"reference":[{"key":"ref_1","first-page":"455","article-title":"17\u03b2-Hydroxysteroid Dehydrogenase 3 Deficiency in the Mediterranean Population","volume":"3","author":"Rosler","year":"2006","journal-title":"Pediatr. Endocrinol. Rev."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"85","DOI":"10.1007\/BF03345572","article-title":"Clinical, Endocrine, and Molecular Findings in 17\u03b2 -Hydroxysteroid Dehydrogenase Type 3 Deficiency","volume":"31","author":"Faienza","year":"2008","journal-title":"J. Endocrinol. Investig."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"229","DOI":"10.1159\/000318004","article-title":"The Clinical and Molecular Heterogeneity of 17\u03b2HSD-3 Enzyme Deficiency","volume":"74","author":"George","year":"2010","journal-title":"Horm. Res. Paediatr."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"666","DOI":"10.1007\/BF03345738","article-title":"17\u03b2-Hydroxysteroid Dehydrogenase-3 Deficiency: From Pregnancy to Adolescence","volume":"32","author":"Bertelloni","year":"2009","journal-title":"J. Endocrinol. Investig."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"650","DOI":"10.1007\/s12020-020-02327-z","article-title":"The Novel Founder Homozygous V225M Mutation in the HSD17B3 Gene Causes Aberrant Splicing and XY-DSD","volume":"69","author":"Zeligson","year":"2020","journal-title":"Endocrine"},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"533","DOI":"10.1590\/S0004-27302012000800012","article-title":"Clinical and Molecular Spectrum of Patients with 17\u03b2-Hydroxysteroid Dehydrogenase Type 3 (17-\u03b2-HSD3) Deficiency","volume":"56","author":"Calais","year":"2012","journal-title":"Arq. Bras. Endocrinol. Metab."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"890","DOI":"10.1097\/AOG.0b013e31824fb482","article-title":"Genome-Wide Fetal Aneuploidy Detection by Maternal Plasma DNA Sequencing","volume":"119","author":"Bianchi","year":"2012","journal-title":"Obstet. Gynecol."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"302","DOI":"10.1002\/uog.17484","article-title":"Analysis of Cell-Free DNA in Maternal Blood in Screening for Aneuploidies: Updated Meta-Analysis: Cell-Free DNA in Screening for Aneuploidies","volume":"50","author":"Gil","year":"2017","journal-title":"Ultrasound Obstet. Gynecol."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"e010002","DOI":"10.1136\/bmjopen-2015-010002","article-title":"Accuracy of Non-Invasive Prenatal Testing Using Cell-Free DNA for Detection of Down, Edwards and Patau Syndromes: A Systematic Review and Meta-Analysis","volume":"6","author":"Freeman","year":"2016","journal-title":"BMJ Open"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"287","DOI":"10.1002\/pd.5632","article-title":"In Case You Missed It: The Prenatal Diagnosis Editors Bring You the Most Significant Advances of 2019","volume":"40","author":"Chitty","year":"2020","journal-title":"Prenat. Diagn."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"656","DOI":"10.1002\/pd.4583","article-title":"Non-invasive Prenatal Diagnosis of Achondroplasia and Thanatophoric Dysplasia: Next-generation Sequencing Allows for a Safer, More Accurate, and Comprehensive Approach","volume":"35","author":"Chitty","year":"2015","journal-title":"Prenat. Diagn."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"439","DOI":"10.1038\/s41591-018-0334-x","article-title":"Non-Invasive Prenatal Sequencing for Multiple Mendelian Monogenic Disorders Using Circulating Cell-Free Fetal DNA","volume":"25","author":"Zhang","year":"2019","journal-title":"Nat. Med."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"950","DOI":"10.1002\/pd.4585","article-title":"Non-Invasive Prenatal Diagnosis for Cystic Fibrosis: Detection of Paternal Mutations, Exploration of Patient Preferences and Cost Analysis","volume":"35","author":"Hill","year":"2015","journal-title":"Prenat. Diagn."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"1026","DOI":"10.1373\/clinchem.2011.178939","article-title":"Digital PCR Analysis of Maternal Plasma for Noninvasive Detection of Sickle Cell Anemia","volume":"58","author":"Barrett","year":"2012","journal-title":"Clin. Chem."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"258","DOI":"10.1002\/pd.4536","article-title":"Non-Invasive Prenatal Diagnostic Testing for \u03b2-Thalassaemia Using Cell-Free Fetal DNA and next Generation Sequencing","volume":"35","author":"Xiong","year":"2015","journal-title":"Prenat. Diagn."},{"doi-asserted-by":"crossref","unstructured":"La Verde, M., De Falco, L., Torella, A., Savarese, G., Savarese, P., Ruggiero, R., Conte, A., Fico, V., Torella, M., and Fico, A. (2021). Performance of Cell-Free DNA Sequencing-Based Non-Invasive Prenatal Testing: Experience on 36,456 Singleton and Multiple Pregnancies. BMC Med. Genom., 14.","key":"ref_16","DOI":"10.1186\/s12920-021-00941-y"},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"1057","DOI":"10.1016\/S0140-6736(01)06183-9","article-title":"Development and Implementation of a New Rapid Aneuploidy Diagnostic Service within the UK National Health Service and Implications for the Future of Prenatal Diagnosis","volume":"358","author":"Mann","year":"2001","journal-title":"Lancet"},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"405","DOI":"10.1038\/gim.2015.30","article-title":"Standards and Guidelines for the Interpretation of Sequence Variants: A Joint Consensus Recommendation of the American College of Medical Genetics and Genomics and the Association for Molecular Pathology","volume":"17","author":"Richards","year":"2015","journal-title":"Genet. Med."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"554","DOI":"10.1136\/adc.2006.098319","article-title":"Consensus Statement on Management of Intersex Disorders","volume":"91","author":"Hughes","year":"2005","journal-title":"Arch. Dis. Child."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"53","DOI":"10.1586\/17446651.4.1.53","article-title":"Diagnosis of 17\u03b2-Hydroxysteroid Dehydrogenase Deficiency","volume":"4","author":"Bertelloni","year":"2009","journal-title":"Expert Rev. Endocrinol. Metab."},{"key":"ref_21","first-page":"4713","article-title":"17\u03b2-Hydroxysteroid Dehydrogenase-3 Deficiency: Diagnosis, Phenotypic Variability, Population Genetics, and Worldwide Distribution of Ancient and de Novo Mutations","volume":"84","author":"Boehmer","year":"1999","journal-title":"J. Clin. Endocrinol. Metab."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"1711","DOI":"10.1007\/s40618-020-01248-y","article-title":"17\u03b2-Hydroxysteroid Dehydrogenase Type 3 Deficiency: Female Sex Assignment and Follow-Up","volume":"43","author":"Faienza","year":"2020","journal-title":"J. Endocrinol. Investig."},{"key":"ref_23","first-page":"130","article-title":"Molecular Genetics and Pathophysiology of 17 Beta-Hydroxysteroid Dehydrogenase 3 Deficiency","volume":"81","author":"Andersson","year":"1996","journal-title":"J. Clin. Endocrinol. Metab."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"20","DOI":"10.1111\/j.1365-2265.2007.02829.x","article-title":"Phenotypic Variability in 17?-Hydroxysteroid Dehydrogenase-3 Deficiency and Diagnostic Pitfalls","volume":"67","author":"Lee","year":"2007","journal-title":"Clin. Endocrinol."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"299","DOI":"10.1097\/00005792-200009000-00003","article-title":"Male Pseudohermaphroditism Due to 17\u03b2-Hydroxysteroid Dehydrogenase 3 Deficiency: Diagnosis, Psychological Evaluation, and Management","volume":"79","author":"Mendonca","year":"2000","journal-title":"Medicine"},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"623","DOI":"10.1515\/jpem-2014-0295","article-title":"Pitfalls in Hormonal Diagnosis of 17-Beta Hydroxysteroid Dehydrogenase III Deficiency","volume":"28","author":"Khattab","year":"2015","journal-title":"J. Pediatric Endocrinol. Metab."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"94","DOI":"10.1016\/j.siny.2017.12.002","article-title":"Beyond Screening for Chromosomal Abnormalities: Advances in Non-Invasive Diagnosis of Single Gene Disorders and Fetal Exome Sequencing","volume":"23","author":"Hayward","year":"2018","journal-title":"Semin. Fetal Neonatal Med."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"424","DOI":"10.1016\/j.ejogrb.2021.01.008","article-title":"Non Invasive Prenatal Testing (NIPT) for Common Aneuploidies and Beyond","volume":"258","author":"Alberry","year":"2021","journal-title":"Eur. J. Obstet. Gynecol. Reprod. Biol."},{"doi-asserted-by":"crossref","unstructured":"Carbone, L., Cariati, F., Sarno, L., Conforti, A., Bagnulo, F., Strina, I., Pastore, L., Maruotti, G.M., and Alviggi, C. (2020). Non-Invasive Prenatal Testing: Current Perspectives and Future Challenges. Genes, 12.","key":"ref_29","DOI":"10.3390\/genes12010015"},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"E333","DOI":"10.1210\/jc.2014-2605","article-title":"Exome Sequencing for the Diagnosis of 46,XY Disorders of Sex Development","volume":"100","author":"Baxter","year":"2015","journal-title":"J. Clin. Endocrinol. Metab."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"243","DOI":"10.1186\/s13059-016-1105-y","article-title":"Disorders of Sex Development: Insights from Targeted Gene Sequencing of a Large International Patient Cohort","volume":"17","author":"Eggers","year":"2016","journal-title":"Genome Biol."}],"container-title":["Genes"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2073-4425\/12\/12\/1890\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T07:35:54Z","timestamp":1760168154000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2073-4425\/12\/12\/1890"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2021,11,25]]},"references-count":31,"journal-issue":{"issue":"12","published-online":{"date-parts":[[2021,12]]}},"alternative-id":["genes12121890"],"URL":"https:\/\/doi.org\/10.3390\/genes12121890","relation":{},"ISSN":["2073-4425"],"issn-type":[{"type":"electronic","value":"2073-4425"}],"subject":[],"published":{"date-parts":[[2021,11,25]]}}}