title: Three-hour analysis of non-invasive foetal sex determination: application of Plexor chemistry.
abstract: BACKGROUND
  The knowledge of the individual genetic "status" in the prenatal era is particularly relevant in the case of positive family history for genetic diseases, in advanced maternal age and in the general screening for foetal abnormalities. In this context, here, we report an innovative molecular assay which utilizes the cell-free foetal DNA (cffDNA) as a source for the early and fast detection of the foetal sex. The study involved 132 pregnant women in their first 3 months of pregnancy, who agreed to give a blood sample. All the collected samples were immediately subjected to the separation of the plasma, which was utilized for the extraction of the cffDNA. Successively, the extracted cffDNA was analysed by a quantitative PCR (qPCR) method based on Plexor-HY chemistry, which is able to simultaneously identify, quantify and discriminate the autosomal DNA from the sex-linked DNA.
  
  
  RESULTS
  Overall, the Plexor-HY assay demonstrated to be sensitive and specific for the determination of low-template DNA, such as the cffDNA. In fact, the Plexor-HY assay has been successfully performed in all the samples, identifying 70 males and 62 females. As the foetal sex can be provided in 120 min just by utilizing a maternal blood sample as cffDNA source, the assay represents a very fast, safe and non-invasive prenatal method.
  
  
  CONCLUSIONS
  The possibility of determining the foetal sex in the early prenatal life consents the application of our assay as a helpful screening test for subjects and families at risk of sex-linked disorders. Moreover, the early knowledge of the foetal sex may be of great help even for the specialist, who might promptly advise the patients concerning the foetal risk of inheriting sex-linked disorders and the clinical utility of performing an invasive prenatal diagnosis.
authors.affiliation: Section of Gynecology and Obstetrics, Academic Department of Biomedicine and Prevention, "Tor Vergata" University Hospital, Oxford Street, 81, 00133, Rome, Italy.|Clinical Department of Surgery, "Tor Vergata" University Hospital, Oxford Street, 81, 00133, Rome, Italy.
authors.forename: Adalgisa
authors.initials: A
authors.lastname: Pietropolli
authors.affiliation: Section of Gynecology and Obstetrics, Academic Department of Biomedicine and Prevention, "Tor Vergata" University Hospital, Oxford Street, 81, 00133, Rome, Italy.|Clinical Department of Surgery, "Tor Vergata" University Hospital, Oxford Street, 81, 00133, Rome, Italy.
authors.forename: Maria Vittoria
authors.initials: MV
authors.lastname: Capogna
authors.affiliation: Department of Biomedicine and Prevention, School of Medicine, University of Rome "Tor Vergata", Montpelier Street, 1, 00133, Rome, Italy. raffaellacascella@virgilio.it.|Emotest Laboratory, M. Licola Patria Street, 60, 80078, Pozzuoli, NA, Italy. raffaellacascella@virgilio.it.
authors.forename: Raffaella
authors.initials: R
authors.lastname: Cascella
authors.affiliation: Molecular Genetics Laboratory UILDM, Santa Lucia Foundation, Ardeatina Street, 306, 00142, Rome, Italy.
authors.forename: Chiara
authors.initials: C
authors.lastname: Germani
authors.affiliation: Section of Gynecology and Obstetrics, Academic Department of Biomedicine and Prevention, "Tor Vergata" University Hospital, Oxford Street, 81, 00133, Rome, Italy.|Clinical Department of Surgery, "Tor Vergata" University Hospital, Oxford Street, 81, 00133, Rome, Italy.
authors.forename: Valentina
authors.initials: V
authors.lastname: Bruno
authors.affiliation: Department of Biomedicine and Prevention, School of Medicine, University of Rome "Tor Vergata", Montpelier Street, 1, 00133, Rome, Italy.
authors.forename: Claudia
authors.initials: C
authors.lastname: Strafella
authors.affiliation: Section of Gynecology and Obstetrics, Academic Department of Biomedicine and Prevention, "Tor Vergata" University Hospital, Oxford Street, 81, 00133, Rome, Italy.|Clinical Department of Surgery, "Tor Vergata" University Hospital, Oxford Street, 81, 00133, Rome, Italy.
authors.forename: Simona
authors.initials: S
authors.lastname: Sarta
authors.affiliation: Section of Gynecology and Obstetrics, Academic Department of Biomedicine and Prevention, "Tor Vergata" University Hospital, Oxford Street, 81, 00133, Rome, Italy.|Clinical Department of Surgery, "Tor Vergata" University Hospital, Oxford Street, 81, 00133, Rome, Italy.
authors.forename: Carlo
authors.initials: C
authors.lastname: Ticconi
authors.affiliation: Department of Biomedicine and Prevention, School of Medicine, University of Rome "Tor Vergata", Montpelier Street, 1, 00133, Rome, Italy.
authors.forename: Giusy
authors.initials: G
authors.lastname: Marmo
authors.affiliation: Department of Biomedicine and Prevention, School of Medicine, University of Rome "Tor Vergata", Montpelier Street, 1, 00133, Rome, Italy.
authors.forename: Sara
authors.initials: S
authors.lastname: Gallaro
authors.affiliation: Department of Biomedicine and Prevention, School of Medicine, University of Rome "Tor Vergata", Montpelier Street, 1, 00133, Rome, Italy.
authors.forename: Giuliana
authors.initials: G
authors.lastname: Longo
authors.affiliation: Department of Biomedicine and Prevention, School of Medicine, University of Rome "Tor Vergata", Montpelier Street, 1, 00133, Rome, Italy.
authors.forename: Luigi Tonino
authors.initials: LT
authors.lastname: Marsella
authors.affiliation: Bambino Gesù Children's Hospital, IRCCS, Sant'Onofrio Square, 4, Rome, Italy.
authors.forename: Antonio
authors.initials: A
authors.lastname: Novelli
authors.affiliation: Department of Biomedicine and Prevention, School of Medicine, University of Rome "Tor Vergata", Montpelier Street, 1, 00133, Rome, Italy.
authors.forename: Giuseppe
authors.initials: G
authors.lastname: Novelli
authors.affiliation: Section of Gynecology and Obstetrics, Academic Department of Biomedicine and Prevention, "Tor Vergata" University Hospital, Oxford Street, 81, 00133, Rome, Italy.|Clinical Department of Surgery, "Tor Vergata" University Hospital, Oxford Street, 81, 00133, Rome, Italy.
authors.forename: Emilio
authors.initials: E
authors.lastname: Piccione
authors.affiliation: Department of Biomedicine and Prevention, School of Medicine, University of Rome "Tor Vergata", Montpelier Street, 1, 00133, Rome, Italy.|Molecular Genetics Laboratory UILDM, Santa Lucia Foundation, Ardeatina Street, 306, 00142, Rome, Italy.
authors.forename: Emiliano
authors.initials: E
authors.lastname: Giardina
mesh_terms: D000328:Adult; D004247:DNA / Q000235:genetics*; D005260:Female; D005333:Fetus; D024282:Genes, sry / Q000235:genetics; D006801:Humans; D008297:Male; D011247:Pregnancy; D011296:Prenatal Diagnosis / Q000379:methods*; D012732:Sex Determination Analysis / Q000379:methods*; D019849:Sex Determination Processes
chemical_list: D004247:DNA