title: Non-invasive pre-implantation aneuploidy screening and diagnosis of beta thalassemia IVSII654 mutation using spent embryo culture medium.
abstract: Cell-free nuclear DNA has been isolated from spent embryo culture medium. Whether this small amount of DNA can be amplified at the whole genome level and the concordance rate of karyotypes and specific alleles between biopsied cells and media has not been evaluated.
  
  
  
  Seven couples were recruited, 88 donated embryos and their corresponding media were collected for whole genome amplification (WGA). The efficiency of WGA, the concordance of chromosome status, and the HBB gene IVSII654 allele between biopsied cells and media were investigated.
  
  
  
  After WGA, the DNA detection rate was 90.90% with a mean concentration of 26.15 ng/μl. The full chromosome concordance rate between biopsied cells and medium was 64.52%, and it increased to 90.00% for diploid blastocyst samples. Analysis of the mutated IVSII654 locus and SNP linkage verified that the DNA present in the medium originated from embryonic cells.
  
  
  
  We confirmed that nuclear DNA is present in spent culture medium and that the majority of this DNA can be amplified for subsequent analysis. Our results showed that non-invasive embryo genetic testing at the chromosomal-level using medium can concordant to the biopsied cells, but it needs further optimized before use in clinical applications. KEY MESSAGES The aggressive biopsy step during PGD/PGS procedure would have a negative effect on the future development of the embryo. Cell-free nuclear DNA has been observed in spent embryo culture medium, which holds promise for the development of non-invasive PGD/PGS approaches. The presence of DNA in medium, its efficiency for WGA, and the concordance between chromosome status and the HBB gene IVSII654 allele as diagnosed from biopsied cells or medium were investigated. Non-invasive embryo genetic testing at the chromosomal-level and allele site using medium can concordant to the biopsied cells, but it needs further optimized before use in clinical applications.
authors: a Department of Clinical Laboratory of Gynecology and Obstetrics, Key Laboratory for Reproduction and Genetics of Guangdong Higher Education Institutes, Key Laboratory for Major Obstetric Diseases of Guangdong Province , Third Affiliated Hospital of Guangzhou Medical University , Guangzhou , PR China.
authors: WeiQiang
authors: W
authors: Liu
authors: b Department of Reproductive Medicine , Third Affiliated Hospital of Guangzhou Medical University , Guangzhou , PR China.
authors: JianQiao
authors: J
authors: Liu
authors: b Department of Reproductive Medicine , Third Affiliated Hospital of Guangzhou Medical University , Guangzhou , PR China.
authors: HongZi
authors: H
authors: Du
authors: b Department of Reproductive Medicine , Third Affiliated Hospital of Guangzhou Medical University , Guangzhou , PR China.
authors: JiaWei
authors: J
authors: Ling
authors: a Department of Clinical Laboratory of Gynecology and Obstetrics, Key Laboratory for Reproduction and Genetics of Guangdong Higher Education Institutes, Key Laboratory for Major Obstetric Diseases of Guangdong Province , Third Affiliated Hospital of Guangzhou Medical University , Guangzhou , PR China.
authors: XiaoFang
authors: X
authors: Sun
authors: a Department of Clinical Laboratory of Gynecology and Obstetrics, Key Laboratory for Reproduction and Genetics of Guangdong Higher Education Institutes, Key Laboratory for Major Obstetric Diseases of Guangdong Province , Third Affiliated Hospital of Guangzhou Medical University , Guangzhou , PR China.
authors: DunJin
authors: D
authors: Chen
mesh_terms: D000782:Aneuploidy; D002474:Cell-Free System; D003470:Culture Media / Q000032:analysis*; D046149:Embryo Culture Techniques / Q000379:methods; D005260:Female; D005820:Genetic Testing / Q000379:methods*; D006454:Hemoglobins / Q000235:genetics*; D006801:Humans; D009154:Mutation; D020641:Polymorphism, Single Nucleotide; D011247:Pregnancy; D017086:beta-Thalassemia / Q000175:diagnosis* / Q000196:embryology / Q000235:genetics
chemical_list: D003470:Culture Media; D006454:Hemoglobins
keywords: Pre-implantation genetic diagnosis and screening; blastocyst; concordance; spent culture medium; whole genome amplification