International Conference on Preimplantation Genetic Diagnosis, İstanbul, Turkey, 8 - 11 May 2013, vol.26, pp.32-33
Conference Paper / Full Text
Akdeniz University Affiliated:
Objective: Preimplantation genetic diagnosis (PGD) for chromosomal
abnormalities has been widely applied for couples
with chromosomal rearrangements, advanced maternal age,
repeated implantation failures, recurrent pregnancy losses and
male infertility. Array comparative genomic hybridization (aCGH)
technique allows simultaneous screening of all chromosomes thus
enabling both aneuploidy testing and detection of segmental
chromosomal abnormalities in the presence of rearrangements.
Here we present an ongoing clinical pregnancy achieved after
PGD using aCGH in a couple with a reciprocal translocation
female and Klinefelter syndrome male.
Materials and methods: An infertile couple, 35-year-old female
partner with a reciprocal translocation 46,XX,t(16;17)(p11;p11)
and male partner with Klinefelter syndrome (47,XXY) with
5 years of primary infertility, with two previous IVF failures,
was referred to the Center for Reproductive Medicine of the
Memorial Antalya Hospital. After informed written consent was
obtained, the female was stimulated by an agonist short protocol
for assisted reproduction and PGD. On the day of oocyte
retrieval, fresh semen sample was taken from male partner
and few motile spermatozoa were used for insemination of
14 mature metaphase II oocytes by intracytoplasmic sperm
injection (ICSI). 13 fertilized zygotes were cultured until day 3.
Twelve embryos with at least 6 blastomeres were selected
for embryo biopsy. One blastomere with a clear nucleus was
removed from each embryo under an inverted microscope.
After enumeration of embryos, corresponding blastomeres were
placed into phosphate buffer saline and transferred to the
genetics facility rapidly at +2 8ºC, maintained on cold blocks.
The genetic analyses were performed in Istanbul Memorial
Hospital Reproductive Genetics Laboratory. DNA extraction and
whole genome amplification (WGA) from blastomeres were
performed with the use of a commercial kit (SurePlex,
BlueGnome, Cambridge, UK). Amplified samples and reference
male DNA were labeled with Cy3 and Cy5, respectively.
Hybridization was performed on 24Sure arrays (BlueGnome,
Cambridge, UK). A dual channel fluorescent laser scanner was used to create images of arrays. Data analysis was performed by
using 24Sure software (BlueGnome, Cambridge, UK).
Results: Results were obtained from all 12 embryos analyzed.
Ten embryos (83.3%) were found to carry abnormalities
associated with the rearrangements, either alone or in combination
with other chromosomal abnormalities. The remaining 2
embryos (16.7%) were either normal or balanced. One normal/
balanced embryo at the morula stage was replaced into the
uterus on day 5. Ten days after embryo transfer, consequent
beta-hCG levels indicated the presence of a pregnancy.
On the 6th gestational week, a clinical pregnancy was confirmed
with a single fetal sac and heart beat. A singleton female ongoing
pregnancy beyond 35 weeks of gestation is present to date.
Conclusion: To the best of our knowledge, this is the first report
presenting an ongoing clinical pregnancy achieved after selection
of chromosomally normal/balanced embryos using the aCGH
technique in a couple where female was a translocation carrier
and male partner was diagnosed with Klinefelter syndrome.
This case report validates the use of aCGH in PGD for
complex indications requiring extensive screening of whole
Keywords: array-comparative genomic hybridization (aCGH),
reciprocal translocation, Klinefelter syndrome.