Assessment of pregnancy rate after comprehensive chromosome screening using next-generation sequencing-based preimplantation genetic diagnosis (NGS PGD) of trophectoderm in frozen blastocyst transfer.

1. Genetic Laboratory
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The impact of next-generation sequencing (NGS)
preimplantation genetic diagnosis (PGD) on pregnancy rate after
frozen blastocyst embryo transfer.
Sebastian Pukszta1, Krzysztof Łukaszuk1,2,3, Celina Cybulska1, Joanna Liss1, Łukasz Płóciennik1, Waldemar
Kuczyński4,5, Lucyna Wójcicka3, Patrycja Kulwikowska1,3, Judyta Zabielska1,3
Study question
Assessment of pregnancy rate after comprehensive chromosome screening using next-generation sequencing-based preimplantation genetic
diagnosis (NGS PGD) of trophectoderm in frozen blastocyst transfer.
1 INVICTA, Fertility and Reproductive Centre, Gdansk, Poland; 2 INVICTA, Fertility and Reproductive Centre, Warsaw, Poland; 3 Department of Nursing, Medical University, Gdansk, Poland;
4 Centre for Reproductive Medicine KRIOBANK, 15-879 Białystok, Poland; 5 Department of Gynecology and Oncological Gynecology, Medical University, 15-276 Białystok, Poland
The trial is registered on www.actrn. org.au with the number: ACTRN12614001037695

2. Genetic Laboratory
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Summary answer
We confirm that NGS can be applied clinically for the purpose of detecting aneuploidy in human preimplantation embryos.
Furthermore, data from this trial indicated that trophectoderm biopsy followed by frozen transfer of euploid embryos was associated
with improvement in implantation and pregnancy rates.
What is known already
Currently used PGD methods of aneuploidy screening have been reported to improve pregnancy rates. However, most of them are
based on low-density array comparative genomic hybridisation (aCGH), with results based on less than 2700 probes. New technical
possibilities, such as next-generation sequencing (NGS) methods, have already shown promise of improved genetic diagnostics in
fresh cycles (1).

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Study design, size, duration
Prospective case control study included analysis of results from 102 embryos obtained from 34 patients (mean age 34) performed between
September 2014 and November 2014 at INVICTA Fertility Centre, Poland.
Participants/materials, setting, methods
Trophectoderm biopsy was performed for the study group embryos. Next generation sequencing with Torrent Suite Software was used for
chromosome copy number variation analysis.

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Fig.1 Examplary result from chip 316.

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Main results and the role of
chance
We transferred 41 vitrified blastocysts during frozen
embryo transfer in the investigated group and 40 in the
control group, resulting in a total of 18 embryos
implanted in the PGD group and 9 in the control group.
The primary outcome measure of clinical pregnancy rate
per transfer was approximately 40 percentage points
higher in the PGD group compared to the control group
(59.3% vs. 20.0%, respectively; p<0.002). The difference
in the implantation rate (secondary outcome measure)
was greater than 20 percentage points (43.9% vs. 22.5%;
p<0.08)
0
10
20
30
40
50
60
70
Clinical Pregnancy Implantation Spontanous abortion
Rate(%)
Frozen embryo transfer
PGD
No PGD
p=0.08
p=0.06
p=0.06

6. Genetic Laboratory
www.invictagenetics.com
INVICTA Genetic Laboratory
ul. Trzy Lipy 3, 80-172 Gdansk, Poland,
T: +48 58 58 58 804, E: pgd@invicta.pl;
W: www.invictagenetics.com
About INVICTA Genetic Laboratory
INVICTA is an experienced genetics laboratory (since 2000)
offering wide range Preimplantation Genetic Diagnosis testing
using state of art Next Generation Sequencing (NGS) techniques.
Wider implications of the findings:
These findings suggest that NGS may be a useful tool for embryo selection that gives normal embryos priority for transfer. NGS
technology allows personalisation of the method and therefore could be used for all cases in which PGD is indicated.
References:
1. Łukaszuk K, Pukszta S, Wells D, et al. Routine use of next-generation sequencing for preimplantation genetic diagnosis of blastomeres obtained from embryos
on day 3 in fresh in vitro fertilization cycles. Fertil Steril. 2015:1-6. doi:10.1016/j.fertnstert.2014.12.123.