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.
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References
Yang Z, Liu J, Collins GS et al: Selection of single blastocysts for fresh transfer via standard morphology assessment alone and with array CGH for good prognosis IVF patients: results from a randomized pilot study. Mol Cytogenet 2012; 5: 24.
Scott Jr RT, Upham KM, Forman EJ et al: Blastocyst biopsy with comprehensive chromosome screening and fresh embryo transfer significantly increases in vitro fertilization implantation and delivery rates: a randomized controlled trial. Fertil Steril 2013; 100: 697–703.
Forman EJ, Hong KH, Ferry KM et al: In vitro fertilization with single euploid blastocyst transfer: a randomized controlled trial. Fertil Steril 2013; 100: 100–107,e1.
Schoolcraft WB, Surrey E, Minjarez D, Gustofson RL, Scott Jr RT, Katz-Jaffe MG : Comprehensive chromosome screening (CCS) with vitrification results in improved clinical outcome in women >35 years: a randomized control trial. Fertil Steril 2012; 98: 718–724.
Bormann CL, Racowsky C : Is universal application of blastocyst biopsy with comprehensive chromosome screening for embryo selection ready for prime time?. Fertil Steril 2013; 2: e5–e6.
Northrop LE, Treff NR, Levy B, Scott Jr RT : SNP microarray-based 24 chromosome aneuploidy screening demonstrates that cleavage-stage FISH poorly predicts aneuploidy in embryos that develop to morphologically normal blastocysts. Mol Hum Reprod 2010; 16: 590–600.
Brezina PR, Benner A, Rechitsky S et al: Single-gene testing combined with single nucleotide polymorphism microarray preimplantation genetic diagnosis for aneuploidy: a novel approach in optimizing pregnancy outcome. Fertil Steril 2011; 95: e5–e8.
Geraedts J, Montag M, Magli MC et al: Polar body array CGH for prediction of the status of the corresponding oocyte. Part I: clinical results. Hum Reprod 2011; 26: 3173–3180.
Scott RT Jr, Ferry K, Su J, Tao X, Scott K, Treff NR : Comprehensive chromosome screening is highly predictive of the reproductive potential of human embryos: a prospective, blinded, nonselection study. Fertil Steril 2012; 97: 870–875.
Treff NR, Su J, Tao X, Levy B, Scott Jr RT : Accurate single cell 24 chromosome aneuploidy screening using whole genome amplification and single nucleotide polymorphism microarrays. Fertil Steril 2010; 94: 2017–2021.
Schoolcraft WB, Treff NR, Stevens JM et al: Live birth outcome with trophectoderm biopsy, blastocyst vitrification, and single-nucleotide polymorphism microarray-based comprehensive chromosome screening in infertile patients. Fertil Steril 2011; 96: 638–640.
Treff NR, Tao X, Ferry KM, Su J, Taylor D, Scott Jr RT : Development and validation of an accurate quantitative real-time polymerase chain reaction-based assay for human blastocyst comprehensive chromosomal aneuploidy screening. Fertil Steril 2012; 97: 819–824, e2.
Mir P, Rodrigo L, Mercader A et al: False positive rate of an arrayCGH platform for single-cell preimplantation genetic screening and subsequent clinical application on day-3. J Assist Reprod Genet 2013; 30: 143–149.
Gutierrez-Mateo C, Colls P, Sanchez-Garcia J et al: Validation of microarray comparative genomic hybridization for comprehensive chromosome analysis of embryos. Fertil Steril 2011; 95: 953–958.
Mastenbroek S, Twisk M, van der Veen F et al: Preimplantation genetic screening: a systematic review and meta-analysis of RCTs. Hum Reprod Update 2011; 17: 454–466.
Capalbo A, Wright G, Elliott T et al: FISH reanalysis of inner cell mass and trophectoderm samples of previously array-CGH screened blastocysts shows high accuracy of diagnosis and no major diagnostic impact of mosaicism at the blastocyst stage. Hum Reprod 2013; 8: 2298–2307.
Gardner DK, Schoolcraft WB . In vitro culture of human blastocysts. Jansen R, Mortimer D . Toward Reproductive Certainty: Fertility and Genetics Beyond. Carnforth, UK: Parthenon Publishing, 1999; 378–388.
Nagy ZP, Chang CC, Shapiro DB et al: Clinical evaluation of the efficiency of an oocyte donation program using egg cryo-banking. Fertil Steril 2008; 2: 520–526.
Capalbo A, Rienzi L, Cimadomo D et al: Correlation between standard blastocyst morphology, euploidy and implantation: an observational study in two centers involving 956 screened blastocysts. Hum Reprod 2014; 6: 1173–1181.
Capalbo A, Bono S, Spizzichino L et al: Sequential comprehensive chromosome analysis on polar bodies, blastomeres and trophoblast: insights into female meiotic errors and chromosomal segregation in the preimplantation window of embryo development. Hum Reprod 2013; 2: 509–518.
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Capalbo, A., Treff, N., Cimadomo, D. et al. Comparison of array comparative genomic hybridization and quantitative real-time PCR-based aneuploidy screening of blastocyst biopsies. Eur J Hum Genet 23, 901–906 (2015). https://doi.org/10.1038/ejhg.2014.222
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DOI: https://doi.org/10.1038/ejhg.2014.222
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