title: Comparison of array comparative genomic hybridization and quantitative real-time PCR-based aneuploidy screening of blastocyst biopsies.
abstract: Comprehensive chromosome screening (CCS) methods are being extensively used to select chromosomally normal embryos in human assisted reproduction. Some concerns related to the stage of analysis and which aneuploidy screening method to use still remain. In this study, the reliability of blastocyst-stage aneuploidy screening and the diagnostic performance of the two mostly used CCS methods (quantitative real-time PCR (qPCR) and array comparative genome hybridization (aCGH)) has been assessed. aCGH aneuploid blastocysts were rebiopsied, blinded, and evaluated by qPCR. Discordant cases were subsequently rebiopsied, blinded, and evaluated by single-nucleotide polymorphism (SNP) array-based CCS. Although 81.7% of embryos showed the same diagnosis when comparing aCGH and qPCR-based CCS, 18.3% (22/120) of embryos gave a discordant result for at least one chromosome. SNP array reanalysis showed that a discordance was reported in ten blastocysts for aCGH, mostly due to false positives, and in four cases for qPCR. The discordant aneuploidy call rate per chromosome was significantly higher for aCGH (5.7%) compared with qPCR (0.6%; P<0.01). To corroborate these findings, 39 embryos were simultaneously biopsied for aCGH and qPCR during blastocyst-stage aneuploidy screening cycles. 35 matched including all 21 euploid embryos. Blinded SNP analysis on rebiopsies of the four embryos matched qPCR. These findings demonstrate the high reliability of diagnosis performed at the blastocyst stage with the use of different CCS methods. However, the application of aCGH can be expected to result in a higher aneuploidy rate than other contemporary methods of CCS.
authors.affiliation: 1] GENERA, Centers for Reproductive Medicine, Rome, Naples, Umbertide, Marostica, Italy [2] GENETYX, Molecular Genetics Laboratory, Marostica, Vicenza, Italy.
authors.forename: Antonio
authors.initials: A
authors.lastname: Capalbo
authors.affiliation: 1] Reproductive Medicine Associates of New Jersey, Basking Ridge, NJ, USA [2] Division of Reproductive Endocrinology, Department of Obstetrics, Gynecology and Reproductive Science, Robert Wood Johnson Medical School, Rutgers University, New Brunswick, NJ, USA.
authors.forename: Nathan R
authors.initials: NR
authors.lastname: Treff
authors.affiliation: 1] GENERA, Centers for Reproductive Medicine, Rome, Naples, Umbertide, Marostica, Italy [2] GENETYX, Molecular Genetics Laboratory, Marostica, Vicenza, Italy.
authors.forename: Danilo
authors.initials: D
authors.lastname: Cimadomo
authors.affiliation: Reproductive Medicine Associates of New Jersey, Basking Ridge, NJ, USA.
authors.forename: Xin
authors.initials: X
authors.lastname: Tao
authors.affiliation: Reproductive Medicine Associates of New Jersey, Basking Ridge, NJ, USA.
authors.forename: Kathleen
authors.initials: K
authors.lastname: Upham
authors.affiliation: 1] GENERA, Centers for Reproductive Medicine, Rome, Naples, Umbertide, Marostica, Italy [2] GENETYX, Molecular Genetics Laboratory, Marostica, Vicenza, Italy.
authors.forename: Filippo Maria
authors.initials: FM
authors.lastname: Ubaldi
authors.affiliation: 1] GENERA, Centers for Reproductive Medicine, Rome, Naples, Umbertide, Marostica, Italy [2] GENETYX, Molecular Genetics Laboratory, Marostica, Vicenza, Italy.
authors.forename: Laura
authors.initials: L
authors.lastname: Rienzi
authors.affiliation: 1] Reproductive Medicine Associates of New Jersey, Basking Ridge, NJ, USA [2] Division of Reproductive Endocrinology, Department of Obstetrics, Gynecology and Reproductive Science, Robert Wood Johnson Medical School, Rutgers University, New Brunswick, NJ, USA.
authors.forename: Richard T
authors.initials: RT
authors.lastname: Scott
mesh_terms: D000782:Aneuploidy; D001755:Blastocyst / Q000166:cytology / Q000378:metabolism*; D025063:Chromosome Disorders / Q000175:diagnosis / Q000235:genetics; D055028:Comparative Genomic Hybridization / Q000379:methods*; D004311:Double-Blind Method; D004622:Embryo, Mammalian / Q000166:cytology / Q000378:metabolism; D005260:Female; D005820:Genetic Testing / Q000379:methods*; D006801:Humans; D008297:Male; D046228:Microarray Analysis / Q000379:methods; D020641:Polymorphism, Single Nucleotide; D011247:Pregnancy; D019836:Preimplantation Diagnosis / Q000379:methods; D011446:Prospective Studies; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity