Single embryo transfer utilizing high resolution or low resolution next generation sequencing for preimplantation genetic testing in ≤35 and >35 year old patients: results from a single center

Abstract Purpose We developed and applied a universal strategy for preimplantation genetic testing for all cystic fibrosis gene mutations (PGT-CF) based on next-generation sequencing (NGS). Methods A molecular protocol was designed to diagnose all CF mutations at preimplantation stage. The detection of CF mutations was performed by direct gene sequencing and linkage strategy testing 38 specific SNPs located upstream and inside the gene for PGT-CF. Seventeen couples at risk of CF transmission decided to undergo PGT-CF. Trophectoderm cell biopsies were performed on day 5–6 blastocysts. PGT for aneuploidy (PGT-A) was performed from the same samples. Tested embryos were transferred on further natural cycles. Results PGT was performed on 109 embryos. Fifteen CF mutations were tested. PGT-CF and PGT-A were conclusive for respectively 92.7% and 95.3% of the samples. A mean of 24.1 SNPs was informative per couple. After a single embryo transfer on natural cycle, 81.3% of the transferred tested embryos were implanted. Conclusions The present protocol based on the entire CFTR gene together with informative SNPs outside and inside the gene can be applied to diagnose all CF mutations at preimplantation stage.

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Effect of next-generation sequencing in preimplantation genetic testing on live birth ratio

Reproduction Fertility and Development ◽

10.1071/rd17428 ◽

2018 ◽

Vol 30 (12) ◽

pp. 1720 ◽

Cited By ~ 2

Author(s):

Joanna Liss ◽

Ewa Pastuszek ◽

Sebastian Pukszta ◽

Eva Hoffmann ◽

Waldemar Kuczynski ◽

...

Keyword(s):

Genetic Testing ◽

Next Generation Sequencing ◽

Embryo Transfer ◽

Live Birth ◽

Live Birth Rate ◽

Frozen Embryo Transfer ◽

Control Group ◽

Next Generation ◽

Preimplantation Genetic Testing ◽

Generation Sequencing

The present study analysed live birth ratios in frozen embryo transfer (FET) cycles where embryo ploidy status was determined with preimplantation genetic testing (PGT) using next-generation sequencing (NGS). PGT was performed on trophectoderm cells biopsied at the blastocyst stage. The present prospective cohort study included 112 women undergoing frozen embryo transfer, with NGS PGT. The control group consisted of 85 patients who underwent the IVF procedure with FET planned for a subsequent cycle. The live birth rate per cycle was higher by ~18.5 percentage points in the investigated compared with control group (42.0% vs 23.5% respectively; P = 0.012). The differences between the study and control groups were also significant for clinical pregnancy (42.0% vs 23.5% respectively; P = 0.012), implantation (41.2% vs 22.2% respectively; P = 0.001) and pregnancy loss rates (9.6% vs 28.6% respectively; P = 0.027). The results show that PGT NGS is a useful method for embryo selection and it may be implemented in routine clinical practice with propitious results.

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Next-Generation Sequencing-Based Pre-Implantation Genetic Testing for Aneuploidy (PGT-A): First Report from Saudi Arabia

Genes ◽

10.3390/genes12040461 ◽

2021 ◽

Vol 12 (4) ◽

pp. 461

Author(s):

Yusra Alyafee ◽

Qamre Alam ◽

Abeer Al Tuwaijri ◽

Muhammad Umair ◽

Shahad Haddad ◽

...

Keyword(s):

Saudi Arabia ◽

Genetic Testing ◽

Next Generation Sequencing ◽

Embryo Transfer ◽

Successful Implementation ◽

Next Generation ◽

Preimplantation Genetic Testing ◽

Reliable Technique ◽

Generation Sequencing

Recently, high-throughput next-generation sequencing (NGS)-based preimplantation genetic testing for aneuploidies techniques came into use. This technique is essential for successful embryo transfer and accomplishing pregnancy, thus reducing the time and cost of additional cycles. In this study, we describe our first experience in introducing an NGS-based preimplantation genetic testing for aneuploidy (PGT-A) service using next-generation sequencing in King Abdulaziz Medical City located in Riyadh, Saudi Arabia. Our main goal was to report the successful implementation of this new technology in clinical practice and highlight the factors that may affect the results. In total, 200 blastomere biopsies were obtained from 36 in vitro fertilization (IVF) cycles from Saudi couples suffering from prolonged infertility or recurrent embryo transfer failure. NGS-based PGT-A was performed in all embryos. The results were analyzed in five age groups, showing that aneuploidy rates increased with maternal age. Moreover, the results also showed that complex abnormal embryos with (2–5) aneuploidy are the most common type of embryos. Additionally, our data showed that chromosome 16-related abnormality was the most frequent abnormality detected among all reported abnormalities. In conclusion, our study suggests that NGS-based PGT-A is an applicable and reliable technique for routine-based embryo screening, especially for couples suffering from recurrent miscarriages or multiple embryo transfer failures.

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