Medium-Based Noninvasive Preimplantation Genetic Diagnosis for Human Α-Thalassemias-sea

AbstractTo develop a noninvasive medium-based preimplantation genetic diagnosis (PGD) test for &agr;-thalassemias-SEA.The embryos of &agr;-thalassemia-SEA carriers undergoing in vitro fertilization (IVF) were cultured. Single cells were biopsied from blastomeres and subjected to fluorescent gap polymerase chain reaction (PCR) analysis; the spent culture media that contained embryo genomic DNA and corresponding blastocysts as verification were subjected to quantitative-PCR (Q-PCR) detection of &agr;-thalassemia-SEA. The diagnosis efficiency and allele dropout (ADO) ratio were calculated, and the cell-free DNA concentration was quantitatively assessed in the culture medium.The diagnosis efficiency of medium-based &agr;-thalassemias–SEA detection significantly increased compared with that of biopsy-based fluorescent gap PCR analysis (88.6% vs 82.1%, P < 0.05). There is no significant difference regarding ADO ratio between them. The optimal time for medium-based &agr;-thalassemias–SEA detection is Day 5 (D5) following IVF.Medium-based &agr;-thalassemias–SEA detection could represent a novel, quick, and noninvasive approach for carriers to undergo IVF and PGD.