title: Development and validation of an accurate quantitative real-time polymerase chain reaction-based assay for human blastocyst comprehensive chromosomal aneuploidy screening.
abstract: OBJECTIVE
  To develop and validate a quantitative real-time polymerase chain reaction (qPCR)-based method for blastocyst trophectoderm comprehensive chromosome screening (CCS) of aneuploidy.
  
  
  METHODS
  Prospective, randomized, and blinded.
  
  
  METHODS
  Academic center for reproductive medicine.
  
  
  METHODS
  Nine cell lines were obtained from a commercial cell line repository, and 71 discarded human blastocysts were obtained from 24 IVF patients that underwent preimplantation genetic screening.
  
  
  METHODS
  None.
  
  
  METHODS
  Consistency of qPCR diagnosis of aneuploidy compared with either conventional karyotyping of cell lines or microarray-based diagnoses of human blastocysts.
  
  
  RESULTS
  Samples from nine cell lines with well characterized karyotypes were diagnosed by qPCR with 97.6% (41/42) consistency. After applying a minimum threshold for concurrence, 100% consistency was achieved. Developmentally normal blastocysts designated as aneuploid or arrested blastocysts designated as euploid by single-nucleotide polymorphism microarray analyses were assigned identical 24 chromosome diagnoses by qPCR in 98.6% of cases (70/71). Overall euploidy (n = 37) and aneuploidy (n = 34) were assigned with 100% consistency. Data was obtained for both sample types in 4 hours.
  
  
  CONCLUSIONS
  These data demonstrate the first qPCR technology capable of accurate aneuploidy screening of all 24 chromosomes in 4 hours. This methodology provides an opportunity to evaluate trophectoderm biopsies with subsequent fresh euploid blastocyst transfer. Randomized controlled trials to investigate the clinical efficacy of qPCR-based CCS are currently underway.
authors.affiliation: Reproductive Medicine Associates of New Jersey, Morristown, New Jersey, USA. ntreff@rmanj.com
authors.forename: Nathan R
authors.initials: NR
authors.lastname: Treff
authors.forename: Xin
authors.initials: X
authors.lastname: Tao
authors.forename: Kathleen M
authors.initials: KM
authors.lastname: Ferry
authors.forename: Jing
authors.initials: J
authors.lastname: Su
authors.forename: Deanne
authors.initials: D
authors.lastname: Taylor
authors.forename: Richard T
authors.initials: RT
authors.lastname: Scott
mesh_terms: D000046:Academic Medical Centers; D000782:Aneuploidy; D001755:Blastocyst / Q000473:pathology*; D002460:Cell Line; D005260:Female; D005307:Fertilization in Vitro / Q000009:adverse effects*; D005820:Genetic Testing; D006801:Humans; D007621:Karyotyping; D009515:New Jersey; D020411:Oligonucleotide Array Sequence Analysis; D011237:Predictive Value of Tests; D011247:Pregnancy; D011248:Pregnancy Complications / Q000175:diagnosis* / Q000235:genetics; D019836:Preimplantation Diagnosis / Q000379:methods*; D011446:Prospective Studies; D060888:Real-Time Polymerase Chain Reaction; D015203:Reproducibility of Results