Medium-Based Noninvasive Preimplantation Genetic Diagnosis for Human α-Thalassemias-SEA

To develop a noninvasive medium-based preimplantation genetic diagnosis (PGD) test for alpha-thalassemias(-SEA). The embryos of alpha-thalassemia(-SEA) carriers undergoing in vitro fertilization (IVF) were cultured. Single cells were biopsied fromblastomeres 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 alpha-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 alpha-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 alpha-thalassemias(-SEA) detection is Day 5 (D5) following IVF. Medium-based alpha-thalassemias(-SEA) detection could represent a novel, quick, and noninvasive approach for carriers to undergo IVF and PGD.