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Medium-based noninvasive preimplantation genetic diagnosis for human α-thalassemias-SEA.

Wu H, Ding C, Shen X, Wang J, Li R, Cai B, Xu Y, Zhong Y, Zhou C - Medicine (Baltimore) (2015)

Bottom Line: The diagnosis efficiency of medium-based α-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.Medium-based α-thalassemias-SEA detection could represent a novel, quick, and noninvasive approach for carriers to undergo IVF and PGD.

View Article: PubMed Central - PubMed

Affiliation: From the Reproductive Medicine Center, First Af&filig;liated Hospital of Sun Yat-sen University, Guangzhou, Guangdong, China, Guangdong Provincial Key Laboratory of Reproductive Medicine.

ABSTRACT
To develop a noninvasive medium-based preimplantation genetic diagnosis (PGD) test for α-thalassemias-SEA. The embryos of α-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 α-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 α-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 α-thalassemias-SEA detection is Day 5 (D5) following IVF. Medium-based α-thalassemias-SEA detection could represent a novel, quick, and noninvasive approach for carriers to undergo IVF and PGD.

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Detection of α-thalassemia-SEA via fluorescent gap PCR analysis. Embryos that had six or more cells early on Day 3 postfertilization were subjected to biopsy and fluorescent gap PCR analysis. A. The top, middle, and lower lanes indicate the normal, affected, and heterozygous, respectively. B. Results summary.
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Figure 1: Detection of α-thalassemia-SEA via fluorescent gap PCR analysis. Embryos that had six or more cells early on Day 3 postfertilization were subjected to biopsy and fluorescent gap PCR analysis. A. The top, middle, and lower lanes indicate the normal, affected, and heterozygous, respectively. B. Results summary.

Mentions: Four hundred thirteen samples from cleavage-stage biopsies were examined for α-thalassemia-SEA via routine fluorescent gap PCR analysis (Figure 1A). There were 108 normal, 103 heterozygous, and 128 affected (α-thalassemia-SEA deletion) embryos, and 74 samples were undetectable. The ratio for the DNA diagnosis efficiency of α-thalassemia-SEA was 82.1% (339/413, Figure 1B).


Medium-based noninvasive preimplantation genetic diagnosis for human α-thalassemias-SEA.

Wu H, Ding C, Shen X, Wang J, Li R, Cai B, Xu Y, Zhong Y, Zhou C - Medicine (Baltimore) (2015)

Detection of α-thalassemia-SEA via fluorescent gap PCR analysis. Embryos that had six or more cells early on Day 3 postfertilization were subjected to biopsy and fluorescent gap PCR analysis. A. The top, middle, and lower lanes indicate the normal, affected, and heterozygous, respectively. B. Results summary.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4554004&req=5

Figure 1: Detection of α-thalassemia-SEA via fluorescent gap PCR analysis. Embryos that had six or more cells early on Day 3 postfertilization were subjected to biopsy and fluorescent gap PCR analysis. A. The top, middle, and lower lanes indicate the normal, affected, and heterozygous, respectively. B. Results summary.
Mentions: Four hundred thirteen samples from cleavage-stage biopsies were examined for α-thalassemia-SEA via routine fluorescent gap PCR analysis (Figure 1A). There were 108 normal, 103 heterozygous, and 128 affected (α-thalassemia-SEA deletion) embryos, and 74 samples were undetectable. The ratio for the DNA diagnosis efficiency of α-thalassemia-SEA was 82.1% (339/413, Figure 1B).

Bottom Line: The diagnosis efficiency of medium-based α-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.Medium-based α-thalassemias-SEA detection could represent a novel, quick, and noninvasive approach for carriers to undergo IVF and PGD.

View Article: PubMed Central - PubMed

Affiliation: From the Reproductive Medicine Center, First Af&filig;liated Hospital of Sun Yat-sen University, Guangzhou, Guangdong, China, Guangdong Provincial Key Laboratory of Reproductive Medicine.

ABSTRACT
To develop a noninvasive medium-based preimplantation genetic diagnosis (PGD) test for α-thalassemias-SEA. The embryos of α-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 α-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 α-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 α-thalassemias-SEA detection is Day 5 (D5) following IVF. Medium-based α-thalassemias-SEA detection could represent a novel, quick, and noninvasive approach for carriers to undergo IVF and PGD.

Show MeSH