Limits...
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.

Show MeSH
Medium-based detection of α-thalassemia-SEA. The affected and undetected embryos determined by biopsy were subsequently cultured in 5 μL G-2 to Day 6. Corresponding medium was collected for detection of α-thalassemia-SEA. The blastocysts were lysed for final confirmation. 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
getmorefigures.php?uid=PMC4554004&req=5

Figure 2: Medium-based detection of α-thalassemia-SEA. The affected and undetected embryos determined by biopsy were subsequently cultured in 5 μL G-2 to Day 6. Corresponding medium was collected for detection of α-thalassemia-SEA. The blastocysts were lysed for final confirmation. A. The top, middle, and lower lanes indicate the normal, affected, and heterozygous, respectively. B. Results summary.

Mentions: As shown in Figure 2A, the VIC S curve along with the flat FAM represents normal Hb (homozygous wild type), whereas the FAM S curve along with the flat VIC represents affected Hb (homozygous mutant). Heterozygous (heterozygous for mutant and wild-type alleles) are shown when both the FAM and VIC indicate S curves.


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)

Medium-based detection of α-thalassemia-SEA. The affected and undetected embryos determined by biopsy were subsequently cultured in 5 μL G-2 to Day 6. Corresponding medium was collected for detection of α-thalassemia-SEA. The blastocysts were lysed for final confirmation. 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 2: Medium-based detection of α-thalassemia-SEA. The affected and undetected embryos determined by biopsy were subsequently cultured in 5 μL G-2 to Day 6. Corresponding medium was collected for detection of α-thalassemia-SEA. The blastocysts were lysed for final confirmation. A. The top, middle, and lower lanes indicate the normal, affected, and heterozygous, respectively. B. Results summary.
Mentions: As shown in Figure 2A, the VIC S curve along with the flat FAM represents normal Hb (homozygous wild type), whereas the FAM S curve along with the flat VIC represents affected Hb (homozygous mutant). Heterozygous (heterozygous for mutant and wild-type alleles) are shown when both the FAM and VIC indicate S curves.

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