title: Blastocoel fluid from differentiated blastocysts harbors embryonic genomic material capable of a whole-genome deoxyribonucleic acid amplification and comprehensive chromosome microarray analysis.
abstract: OBJECTIVE
  To obtain embryonic molecular karyotypes from genomic DNA (deoxyribonucleic acid) isolated from blastocoel fluid (BF) and to compare these karyotypes with the karyotypes from the remaining inner cell mass (ICM) and trophectoderm (TE) of the blastocyst.
  
  
  METHODS
  Prospective cohort study.
  
  
  METHODS
  Academic center and preimplantation genetics laboratory.
  
  
  METHODS
  Ninety-six donated cryopreserved embryos.
  
  
  METHODS
  Embryo biopsy, BF aspiration, DNA analysis using a comparative genomic hybridization microarray (aCGH).
  
  
  METHODS
  The aCGH of a single blastomere, BF-DNA, and ICM-TE.
  
  
  RESULTS
  The BF-DNA samples resulted in a successful aCGH in 63% of cases. Discordance in karyotypes was found between the BF-DNA and the ICM-TE in 52% of cases. A total of 70% of aneusomic (mosaicism), cleavage-stage embryos differentiated into euploid blastocysts. Probabilities for diagnostic accuracy were calculated and demonstrated the following: sensitivity of 0.88 (95% confidence interval [CI]: 0.62-0.98); specificity of 0.55 (95% CI: 0.39-0.70); positive predictive value of 0.41 (95% CI: 0.25-0.60); negative predictive value of 0.92 (95% CI: 0.75-0.99).
  
  
  CONCLUSIONS
  Genomic DNA from the BF can be amplified and characterized by comprehensive chromosome microarrays. The results demonstrated that aneusomic cleavage-stage embryos differentiated into euploid blastocysts, possibly using a mechanism that marginalizes aneuploid nuclei into the blastocoel cavity. In addition, owing to the high discordance between the karyotypes obtained from the BF-DNA and the ICM-TE, using BF-DNA for preimplantation genetic testing is not yet advised.
authors.affiliation: Division of Reproductive Endocrinology and Infertility, Department of Gynecology and Obstetrics, Johns Hopkins University School of Medicine, Baltimore, Maryland; Department of Obstetrics and Gynecology, Womack Army Medical Center, Fort Bragg, North Carolina. Electronic address: kyle.tobler@gmail.com.
authors.forename: Kyle J
authors.initials: KJ
authors.lastname: Tobler
authors.affiliation: Division of Reproductive Endocrinology and Infertility, Department of Gynecology and Obstetrics, Johns Hopkins University School of Medicine, Baltimore, Maryland.
authors.forename: Yulian
authors.initials: Y
authors.lastname: Zhao
authors.affiliation: Fort Worth Fertility, Fort Worth, Texas.
authors.forename: Ric
authors.initials: R
authors.lastname: Ross
authors.affiliation: AdvaGenix, LLC, Rockville, Maryland.
authors.forename: Andy T
authors.initials: AT
authors.lastname: Benner
authors.affiliation: AdvaGenix, LLC, Rockville, Maryland.
authors.forename: Xin
authors.initials: X
authors.lastname: Xu
authors.affiliation: AdvaGenix, LLC, Rockville, Maryland.
authors.forename: Luke
authors.initials: L
authors.lastname: Du
authors.affiliation: Division of Reproductive Endocrinology and Infertility, Department of Gynecology and Obstetrics, Johns Hopkins University School of Medicine, Baltimore, Maryland.
authors.forename: Kathleen
authors.initials: K
authors.lastname: Broman
authors.affiliation: Division of Reproductive Endocrinology and Infertility, Department of Gynecology and Obstetrics, Johns Hopkins University School of Medicine, Baltimore, Maryland.
authors.forename: Kim
authors.initials: K
authors.lastname: Thrift
authors.affiliation: Division of Reproductive Endocrinology and Infertility, Department of Gynecology and Obstetrics, Vanderbilt University School of Medicine, Memphis, Tennessee; Department of Surgery, St. Jude Children's Research Hospital, Memphis, Tennessee; Fertility Associates of Memphis, Memphis, Tennessee.
authors.forename: Paul R
authors.initials: PR
authors.lastname: Brezina
authors.affiliation: Division of Reproductive Endocrinology and Infertility, Department of Gynecology and Obstetrics, Johns Hopkins University School of Medicine, Baltimore, Maryland; AdvaGenix, LLC, Rockville, Maryland.
authors.forename: William G
authors.initials: WG
authors.lastname: Kearns
mesh_terms: D000328:Adult; D001755:Blastocyst / Q000502:physiology*; D015331:Cohort Studies; D055028:Comparative Genomic Hybridization / Q000379:methods*; D005260:Female; D006801:Humans; D046228:Microarray Analysis / Q000379:methods*; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*; D011247:Pregnancy; D011446:Prospective Studies
keywords: Blastocoel fluid; comparative genomic hybridization; microarray; preimplantation genetic screening; preimplantation genetic testing