DEVELOPMENT AND CLINICAL APPLICATION OF A PREIMPLANTATION GENETIC TESTING FOR MONOGENIC DISEASE (PGT-M) FOR BETA THALASSEMIA IN VIETNAM

Đào Mai Anh, Gary L Harton, Nguyễn Quang Vinh, Nguyễn Văn Huynh, Hoàng Thị Nhung, Phạm Thúy Nga, Lê Thị Thu Hiền, Nguyễn Minh Đức, Trần Quốc Quân

Main Article Content

Abstract

Purpose: The purpose of this research is to study the clinical outcomes using a next-generation sequencing-based protocol allowing for simultaneous testing of mutations in the beta thalassemia (HBB) gene, including single nucleotide polymorphism (SNP) markers for PGT-M along with low-pass whole genome analysis of chromosome aneuploidies for PGT-A. Methods: A combined PGT-M (thalassemia) plus PGT-A system was developed for patients undergoing IVF in Vietnam. Here we developed a system for testing numerous thalassemia mutations plus SNP-based testing for backup mutation analysis and contamination control using next-generation sequencing (NGS). Low -pass next-generation sequencing was used to assess aneuploidy in some of the clinical PGT cases. Patients underwent IVF followed by embryo biopsy at the blastocyst stage for combined PGT-A/M. Results: Two cases have completed the entire process including transfer of embryos, while a further nine cases have completed the IVF and PGT-M/A analysis but have not completed embryo transfer. In the two cases with embryo transfer, both patients achieved pregnancy with an unaffected, euploid embryo confirmed through prenatal diagnosis. In the further nine cases, 39 embryos were biopsied and all passed QC for amplification. There were 8 unaffected embryos, 31 carrier embryos, and 11 affected embryos. A subset of 24 embryos also had PGT-A analysis with 22 euploid embryos and 2 aneuploid embryos. Conclusions: Here we report the development and clinical application of a combined PGT-M for HBB and PGT-A for gross chromosome aneuploidies from 11 patients with detailed laboratory findings along with 2 cases that have completed embryo transfer.

Article Details

References

1. Agilent-OnePGT Library Prep for Illumina Sequencers. https:// www.agilent.com/ cs/library/ usermanuals/ public/ G9425-90000. pdf. Accessed Nov 4 2020.
2. De Rycke M, Goossens V, Kokkali G, Meijer-Hoogeveen M, Coonen E, Moutou C.: “EHSRE PGD consortium data collection XIV-XV: cycles from January 2011 to December 2012 with pregnancy follow-up to October 2013”. Hum Reprod. 2017; 32(10):1974–94.
3. Dreesen J, Destouni A, Kourlaba G, Degn B, Mette WC, Carvalho F, et al.: “Evaluation of PCR-based preimplantation genetic diagnosis applied to monogenic diseases: a collaborative ESHRE PGD consortium study”. Eur J Hum Genet. 2014; 22(8):1012–8.
4. Findlay I, Ray P, Quirke P, Rutherford A, Lilford R.: “Allelic dropout and preferential amplification in single cells and human blastomeres: implications for preimplantation diagnosis of sex and cystic fibrosis”. Hum Reprod. 1995;10(6):1609–18.
5. Forman EJ, Hong KH, Ferry KM, Tao X, Taylor D, Levy B, Treff NR, Scott Jr RT.: ”In vitro fertilization with single euploid blastocyst transfer: a randomized controlled trial”. Fertil Steril. 2013; 100(1):100–7.
6. Franasiak JM, Forman EJ, Hong KH, Werner MD, Upham KM, Treff NR, et al.: ”The nature of aneuploidy with increasing age of the female partner: a review of 15,169 consecutive trophectoderm biopsies evaluated with comprehensive chromosomal screening”. Fertil Steril. 2014;101(3):656–63.
7. Galanello R, Origa R.: “Beta-thalassemia”. Orphanet J Rare Dis. 2010;5:11.
8. Và nhiều tài liệu tiếng nước ngoài khác.