Detection of SEA-type α-thalassemia in embryo biopsies by digital PCR

Lee, Ta-Hsien; Hsu, Ya-Chiung; Chang, Chia‐Lin

doi:10.1016/j.tjog.2017.03.002

Cited by 10 publications

(3 citation statements)

References 20 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…In this study, we presented a comprehensive PGT approach for SEA-type α-thalassemia by fluorescent Gap-PCR combined with SNP array, which can directly detect SEA-type α-thalassemia along with linkage analysis as well as aneuploidy screening. The detection methods of SEA-type α-thalassemia PGT evolved from early PCR technologies (Deng et al, 2006;Wirawit et al, 2012;Ta-Hsien et al, 2017), to STR polymorphic markers (Aspasia et al, 2012), then to recent SNP haplotype analysis based on NGS (Chen et al, 2017;Chen et al, 2020;Li et al, 2020). Direct detection of target genes potentially leads to misdiagnosis due to ADO.…”

Section: Discussionmentioning

confidence: 99%

A comprehensive preimplantation genetic testing approach for SEA-type α-thalassemia by fluorescent gap-polymerase chain reaction combined with haplotype analysis

Wang,

Ma,

Guo

et al. 2023

Front. Genet.

View full text Add to dashboard Cite

Introduction: This study aimed to evaluate the feasibility and necessity of using fluorescence Gap-polymerase chain reaction combined with haplotype analysis in preimplantation genetic testing for SEA-type α-thalassemia.Methods: A total of 26 preimplantation genetic testing biopsy cycles were performed in 25 families from June 2021 to February 2022. All couples were carriers of SEA-type α-thalassemia. Fluorescent Gap-polymerase chain reaction was used for detecting fragment deletion. Subsequently, according to the results of polymerase chain reaction, reference embryos were identified to establish haplotype using single nucleotide polymorphism array, and aneuploidy was screened simultaneously. In cases wherein the polymerase chain reaction results were inconsistent with the haplotype results, the reasons were investigated, either by retest of the biopsied samples or rebiopsy of the embryo.Results: Among the 172 embryos, 162 had consistent results when tested using both methods, resulting in a consistency rate of 94.2%. Conversely, 10 embryos had inconsistent results, mainly due to chromosome 16 aneuploidy (n = 7), allele dropout in Gap-polymerase chain reaction (n = 2), or incorrect haplotype due to poor sample amplification quality (n = 1). The clinical pregnancy rate of each frozen-thawed embryo transfer was 57.7% (15/26). Six families underwent prenatal diagnosis, which confirmed the results of preimplantation genetic testing.Conclusion: Fluorescent Gap-polymerase chain reaction combined with haplotype analysis is feasible and necessary for SEA-type α-thalassemia preimplantation genetic testing.

show abstract

Section: Discussionmentioning

confidence: 99%

A comprehensive preimplantation genetic testing approach for SEA-type α-thalassemia by fluorescent gap-polymerase chain reaction combined with haplotype analysis

Wang,

Ma,

Guo

et al. 2023

Front. Genet.

View full text Add to dashboard Cite

show abstract

“…To identify normal a-globin and SEA-type deletion alleles, WGA products from all biopsy samples were amplified with a nested PCR protocol and second PCR reaction. The PCR protocol and primers were carried out in accordance with a previous study [ 14 ].…”

Section: Methodsmentioning

confidence: 99%

Using affected embryos to establish linkage phase in preimplantation genetic testing for thalassemia

Deng

Liang

et al. 2022

Reprod Biol Endocrinol

View full text Add to dashboard Cite

Background This study aimed to evaluate the ability of next-generation sequencing (NGS) to conduct preimplantation genetic testing (PGT) for thalassemia using affected embryos. Methods This study included data from 36 couples who underwent PGT for thalassemia without probands and relative pedigrees. NGS results were compared with prenatal diagnosis results. Results Thirty-six couples (29 α-thalassemia and 7 β-thalassemia) underwent 41 PGT cycles (31 α-thalassemia and 10 β-thalassemia). Analysis using NGS produced conclusive results for all biopsied blastocysts (100%, 217/217). One hundred and sixty (73.7%, 160/217) were unaffected by thalassemia. Preimplantation genetic testing for aneuploidy revealed that 112 (70.0%, 112/160) were euploid. Single blastocysts were transferred into the uteri of 34 women (53 frozen embryo transfer [FET] cycles). Thirty-two cycles resulted in clinical pregnancies, with a clinical pregnancy rate of 60.1% (32/53) per FET cycle. Twenty-two cycles (22 couples) resulted in 23 live births, with a live birth rate of 43.4% (23/53; 3 cycles were ongoing pregnancies). All 25 embryos’ prenatal diagnosis results and/or thalassemia gene analyses after delivery were concordant with the NGS-PGT results. Seven embryos (21.9%, 7/32) were miscarried before 12 weeks’ gestation, and the abortion villus in four showed a normal karyotype and thalassemia results consistent with the NGS-PGT results. Aborted fetus samples from 3 cycles were not available because the pregnancy lasted less than 5 weeks. Conclusion NGS can be used to conduct PGT for thalassemia using affected embryos as a reference. Trial registration Retrospectively registered.

show abstract

“…伴随分子生物学技术的发展, 数字PCR、高分辨率溶解曲线分析、探针等技术开始用于地贫的人群筛查及产前诊断, 极大地提高了精确度及诊断速度, 同时降低了DNA量的需求 [16,17] , 最终通过优生指导, 有效降低重型地贫患儿的出生率, 从根本上减轻家庭与社会的压力和负担.…”

Section: 通过Crispr-cas9技术去除β型地贫患者hsc细胞中unclassified

Research progress of hereditary blood diseases

Li¹,

Yang²

2017

Sci. Sin.-Vitae

View full text Add to dashboard Cite

Applications of proteomics in hepatic diseases research Science in China Series C-Life Sciences 47, 101 (2004); Roles and mechanisms of ginsenoside in cardiovascular diseases: progress and perspectives SCIENCE CHINA Life Sciences 59, 292 (2016); Advances in research into gamete and embryo-fetal origins of adult diseases SCIENCE CHINA Life Sciences 62, 360 (2019); Platelet proteomics and its advanced application for research of blood stasis syndrome and activated blood circulation herbs of Chinese medicine

show abstract

Detection of SEA-type α-thalassemia in embryo biopsies by digital PCR

Cited by 10 publications

References 20 publications

A comprehensive preimplantation genetic testing approach for SEA-type α-thalassemia by fluorescent gap-polymerase chain reaction combined with haplotype analysis

A comprehensive preimplantation genetic testing approach for SEA-type α-thalassemia by fluorescent gap-polymerase chain reaction combined with haplotype analysis

Using affected embryos to establish linkage phase in preimplantation genetic testing for thalassemia

Research progress of hereditary blood diseases

Contact Info

Product

Resources

About