Preimplantation genetic diagnosis by fluorescence in situ hybridization of reciprocal and Robertsonian translocations.

OBJECTIVE: The presence of reciprocal and Robertsonian chromosomal rearrangement is often related to recurrent miscarriage. Using preimplantation genetic diagnosis, the abortion rate can be decreased. Cases treated at our center were reviewed. MATERIALS AND METHODS: A retrospective analysis for either Robertsonian or reciprocal translocations was performed on all completed cycles of preimplantation genetic diagnosis at our center since the first reported case in 2004 until the end of 2010. Day 3 embryo biopsies were carried out, and the biopsied cell was checked by fluorescent in situ hybridization using relevant informative probes. Embryos with a normal or balanced translocation karyotype were transferred on Day 4. RESULTS: Thirty-eight preimplantation genetic diagnosis cycles involving 17 couples were completed. A total of 450 (82.6%) of the total oocytes were MII oocytes, and 158 (60.0%) of the two-pronuclei embryos were biopsied. In 41.4% of the fluorescent in situ hybridization analyses, the results were either normal or balanced. Embryos were transferred back after 21 cycles. Three babies were born from Robertsonian translocation carriers and another two from reciprocal translocation carriers. The miscarriage rate was 0%. Among the reciprocal translocation group, the live delivery rate was 8.3% per ovum pick-up cycle and 18.2% per embryo transfer cycle. Among the Robertsonian translocation group, the live delivery rate was 14.3% per ovum pick-up cycle and 20.0% per embryo transfer cycle. CONCLUSION: There is a trend whereby the outcome for Robertsonian translocation group carriers is better than that for reciprocal translocation group carriers. Aneuploidy screening may possibly be added in order to improve the outcome, especially for individuals with an advanced maternal age. The emergence of an array-based technology should help improve this type of analysis.

Low-dose GnRH antagonist protocol is as effective as the long GnRH agonist protocol in unselected patients undergoing in vitro fertilization and embryo transfer.

OBJECTIVE: The present retrospective and controlled comparative study was designed to evaluate the pregnancy rate achieved using a modified, fixed, multiple-dose 0.125mg gonadotropin-releasing hormone (GnRH) antagonist protocol with the long GnRH agonist protocol as the control group. MATERIALS AND METHODS: One hundred and twenty unselected women between 30 and 40 years of age, in their first cycle of IVF/ICSI, with a baseline follicle-stimulating hormone (FSH) <10 IU and an antral follicle count >3 were assigned into two groups: (1) the study group received 0.125mg of cetrorelix daily starting on Day 6 of stimulation; and (2) the control group received leuprolide daily starting in the mid-luteal phase of the preceding cycle. Both groups were given a flexible dose of recombinant FSH for stimulation. An ongoing pregnancy rate of more than 12 weeks was the primary outcome measure of the study. RESULTS: Primary and secondary outcomes were comparable in both groups. A shorter duration of stimulation, a lower dosage of recombinant FSH consumption and a thinner endometrium on the day of human chorionic gonadotropin administration were all observed in the GnRH antagonist group. CONCLUSION: A dosage of 0.125mg GnRH antagonist protocol was effective for these unselected patients during IVF/ET.

The relationship of pronuclear stage morphology and chromosome status at cleavage stage.

PURPOSE: Infertile couples undergoing in vitro fertilization-embryo transfer (IVF-ET) program were included to study the relationship of pronuclear stage morphology and chromosome status at cleavage stage. METHODS: Eighteen to twenty-one hours after fertilization, zygotes were checked for pronuclear morphology with modified Scott Z-score system. After embryo transfer on day 3, arrested or non-transferred 2 PN embryos were spread for fluorescence in situ hybridization (FISH) staining of probes to chromosomes 18, X and Y. RESULTS: Ninety-eight embryos were successfully fixed and stained. The chromosome status were recorded in each 2 PN score group: 7 (54%) of 13 embryos in Z2 group, 14 (35%) of 40 in Z3 group and 10 (36%) of 28 in Z4 group being normal diploid. Z1 group has 12 (71%) of 17 embryos being normal diploid, which is significantly more than Z3 (p = 0.020) and Z4 group (p = 0.033). CONCLUSION: Our results demonstrated a high probability to get normal diploid embryos if good morphology at pronuclear stage was used as selection criteria, especially for Z1 score embryos.

Pre-implantation genetic diagnosis: a preliminary report of 2 years of experience.

BACKGROUND: Pre-implantation genetic diagnosis (PGD) is defined as making a diagnosis or screening embryos or gametes before implantation. It has the advantage of avoiding repeated spontaneous abortions or therapeutic termination of pregnancy resulting from abnormal embryos. Here, we present our preliminary report of 2 years of experience. METHODS: From March 2001 through October 2002, couples seeking assistance for in vitro fertilization (IVF) were referred for PGD due to chromosomal problems or for aneuploidy screening (PGD-AS). One or two blastomeres were aspirated on day 3 and analyzed using the fluorescent in situ hybridization (FISH) technique. Probes to chromosomes X, Y, and 18 were used for aneuploidy screening and individual specific probes were chosen for chromosomal translocations. Unaffected embryos were transferred on day 5. RESULTS: There were 25 cycles for aneuploid screening (group 1) and four cycles for chromosomal translocation (group 2). In group 1, 73 embryos were biopsied with a successful biopsy/fixation rate of 72.6% and a diagnosis rate of 96.2%. Fifteen unaffected embryos were transferred in 11 cycles, achieving two sets of twins and four singleton pregnancies (implantation rate: 53.3%). In group 2, 27 embryos were biopsied with a successful biopsy/fixation rate of 66.7% and a diagnosis rate of 88.9%. Seven non-affected embryos were transferred in three cycles, resulting in one set of twins (implantation rate: 33.3%). All antenatal amniocentesis confirmed the diagnosis. Post-natal physical examination showed no evidence of major abnormalities. CONCLUSIONS: PGD is an alternative method for having healthy children in selected couples with chromosomal abnormalities. In addition, PGD-AS may increase the implantation rate in infertile couples seeking IVF assistance.