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Development and validation of an accurate quantitative real time
1. Development and validation of an accurate
quantitative real-time polymerase chain reactionâ
based assay for human blastocyst comprehensive
chromosomal aneuploidy screening
Author : Nathan R. Treff (Ph.D), et al.
Fertility and Sterility (IF: 4.174) VOL. 97 NO. 4 / APRIL 2012
Speaker : Hung,
Mau-Ren
1
3. The Reason We Start
1. Enhance embryo selection
2. Increase implantation rates
3. Reduce the incidence of miscarriage
4. Reduce the time of implantation
3
16. Two-phase design
Phase I
Using the cell-lines to evaluate
the system and establish
baseline database.
Phase II
Use 71 of blastocysts to
evaluate the system.
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20. Statistics
ÎCt was calculated from the average âCt of the
16 reactions targeting a specific Chromosome
minus the average â Ct of all of the 336 reactions
targeting all of the remaining autosomes
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21. Statistics
ÎCt was calculated from
the average âCt of the 16
reactions targeting a
specific Chromosome
minus the average â Ct of
all of the 336 reactions
targeting all of the
remaining autosomes
4
16
Specific Chromosome
23
22. Statistics
ÎCt was calculated from
the average âCt of the 16
ÎCt a 4
reactions targeting
specific Chromosome
minus the average â Ct of
all of the 336 reactions
targeting all of the
remaining autosomes
16
4
21
336
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25. FIGURE 1
Examples of qPCR-based
24-chromosome copy
number results from 5cell samples derived from
nine cell lines with
previously well
characterized karyotypes.
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28. FIGURE 2
97.6% reliability of obtaining a
diagnosis and a100% level of
consistency of chromosome-specific
(n =984) and 24-chromosome copy
number (n =41) assignments
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33. Back to the start âŠ
1. Enhance embryo selection
2. Increase implantation rates
PGD
3. Reduce the incidence of miscarriage
4. Reduce the time of implantation
5. Cost-Down
qPCR
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