title: Potential of syncytiotrophoblasts isolated from the cervical mucus for early non-invasive prenatal diagnosis: evidence of a vanishing twin.
abstract: BACKGROUND
  Non-invasive methods to assess the foetal genome during pregnancy will provide new opportunities to offer pregnant women a more comprehensive genetic diagnosis of their established foetus. The aim of this study was to determine the presence and frequency of foetal cells in transcervical cell (TCC) mucus samples from pregnant women and determine their suitability for early prenatal diagnosis.
  
  
  METHODS
  Syncytiotrophoblasts in aspirated TCC mucus samples were identified by immunostaining with the foetal-specific antibody NDOG1. Genetic analysis of foetal cells was performed by laser capture microdissection and quantitative fluorescent PCR (QF-PCR).
  
  
  RESULTS
  In 116 of 207 (56%) TCC samples, abundant syncytiotrophoblasts were retrieved. However, when TCC samples were stratified for the presence of chorionic villous fragments, syncytiotrophoblasts were identified in 85 of 109 (78%) samples. Significant numbers of syncytiotrophoblasts were found in TCC samples collected between 6 and 9weeks of gestation (mean 741, range 25-2884). QF-PCR analysis of NDOG1 positive syncytiotrophoblasts and matching maternal DNA confirmed their foetal origin and correct foetal cell sexing was achieved in 97% of TCC samples. The one discordant sex diagnosis was associated with a dizygotic dichorionic twin pregnancy resulting from the implantation of a female T21 embryo and a normal male embryo, where the female T21 foetus had succumbed at 6weeks of gestation and was vanishing.
  
  
  CONCLUSIONS
  Syncytiotrophoblasts can be successfully isolated from TCC samples and represent a suitable source of cells for genetic analysis of the established foetus in early pregnancy. The study highlights a vanishing twin as a potential cause for discordant non-invasive prenatal test results.
authors.affiliation: Genetic Technologies Ltd, Melbourne, Australia.
authors.forename: Debbie
authors.initials: D
authors.lastname: Mantzaris
authors.affiliation: Department of Anatomy and Developmental Biology, Monash University, Melbourne, Australia; Berry Genomics, Beijing, China. Electronic address: david.cram@monash.edu.
authors.forename: David S
authors.initials: DS
authors.lastname: Cram
mesh_terms: D037881:Aborted Fetus; D000328:Adult; D002582:Cervix Mucus / Q000166:cytology*; D005260:Female; D005333:Fetus / Q000737:chemistry* / Q000473:pathology; D005865:Gestational Age; D006801:Humans; D008297:Male; D011247:Pregnancy; D011261:Pregnancy Trimester, First; D011296:Prenatal Diagnosis / Q000379:methods*; D012732:Sex Determination Analysis / Q000379:methods*; D014327:Trophoblasts / Q000378:metabolism* / Q000473:pathology; D014429:Twins, Dizygotic
keywords: Down syndrome; Laser capture microdissection; Non-invasive prenatal diagnosis; Quantitative fluorescent-PCR; Transcervical cell sampling; Vanishing twin