Development of pre-implantation genetic testing protocol for monogenic disorders (PGT-M) of Hb H disease.

Autor: Somboonchai P; Department of Obstetrics and Gynaecology, Faculty of Medicine, Chiang Mai University, 110 Intawaroros Road, Sripoom, Mueang, Chiang Mai, 50200, Thailand., Charoenkwan P; Department of Pediatrics, Faculty of Medicine, Chiang Mai University, 110 Intawaroros Road, Sripoom, Mueang, Chiang Mai, 50200, Thailand., Piyamongkol S; Department of Pharmaceutical Sciences, Faculty of Pharmacy, Chiang Mai University, 239 Suthep Road, Suthep, Mueang, Chiang Mai, 50200, Thailand., Lattiwongsakorn W; Department of Obstetrics and Gynaecology, Faculty of Medicine, Chiang Mai University, 110 Intawaroros Road, Sripoom, Mueang, Chiang Mai, 50200, Thailand., Pantasri T; Department of Obstetrics and Gynaecology, Faculty of Medicine, Chiang Mai University, 110 Intawaroros Road, Sripoom, Mueang, Chiang Mai, 50200, Thailand. tawiwan.p@cmu.ac.th., Piyamongkol W; Department of Obstetrics and Gynaecology, Faculty of Medicine, Chiang Mai University, 110 Intawaroros Road, Sripoom, Mueang, Chiang Mai, 50200, Thailand. wirawit.p@cmu.ac.th.
Jazyk: angličtina
Zdroj: BMC genomics [BMC Genomics] 2024 Jul 03; Vol. 25 (1), pp. 668. Date of Electronic Publication: 2024 Jul 03.
DOI: 10.1186/s12864-024-10578-7
Abstrakt: Hb H disease is the most severe form of α-thalassemia compatible with post-natal life. Compound heterozygous α 0 -thalassemia - SEA deletion/α + -thalassemia - 3.7kb deletion is the commonest cause of Hb H disease in Thailand. Preimplantation genetics testing for monogenic disorders (PGT-M) is an alternative for couples at risk of the disorder to begin a pregnancy with a healthy baby. This study aims to develop a novel PCR protocol for PGT-M of Hb H disease - SEA/-3.7kb using multiplex fluorescent PCR. A novel set of primers for α + -thalassemia - 3.7kb deletion was developed and tested. The PCR protocol for α 0 -thalassemia - SEA deletion was combined for Hb H disease - SEA/-3.7kb genotyping. The PCR protocols were applied to genomic DNA extracted from subjects with different thalassemia genotypes and on whole genome amplification (WGA) products from clinical PGT-M cycles of the families at risk of Hb Bart's. The results were compared and discussed. The results showed three PCR products from α + -thalassemia - 3.7kb primer set, and three from α 0 thalassemia SEA primer set. The results were consistent with the known thalassemia genotypes. The novel -α 3.7 primers protocol was also tested on 37 WGA products from clinical PGT-M cycles giving accurate genotyping results and a satisfying amplification efficiency with the ADO rates of 2.7%, 0%, and 0% for HBA2, HBA1, and internal control fragments, respectively. This novel PCR protocol can precisely distinguish Hb H disease - SEA/-3.7kb from other genotypes. Additionally, this is the first PCR protocol for Hb H disease - SEA/-3.7kb which is optimal for PGT-M.
(© 2024. The Author(s).)
Databáze: MEDLINE
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