The successful strategy of comprehensive pre-implantation genetic testing for beta-thalassaemia–haemoglobin E disease and chromosome balance using karyomapping

Piyamongkol, Sirivipa; Mongkolchaipak, Suchada; Charoenkwan, Pimlak; Sirapat, Rungthiwa; Suriya, Wanwisa; Pantasri, Tawiwan; Tongsong, Theera; Piyamongkol, Wirawit

doi:10.1080/01443615.2022.2070728

Cited by 2 publications

(5 citation statements)

References 21 publications

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“…Currently, PGT offers a crucial alternative strategy to avoid the transmission of pathogenic variants for couples diagnosed with rare monogenic diseases through carrier screening or other genetic diagnoses ( Xi et al, 2020 ; Yang et al, 2022a ; Bai et al, 2022 ; Piyamongkol et al, 2022b ; Yang et al, 2022b ; Zhang et al, 2022 ; Xu et al, 2023 ). In mainland China, as of December 2022, there are 93 institutions that can do PGT, which can basically cover all the Chinese people.…”

Section: Discussionmentioning

confidence: 99%

Integrative preimplantation genetic testing analysis for a Chinese family with hereditary spherocytosis caused by a novel splicing variant of SPTB

Tian,

Wang,

Yang

et al. 2023

Front. Genet.

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Hereditary spherocytosis (HS), the most common inherited hemolytic anemia disorder, is characterized by osmotically fragile microspherocytic red cells with a reduced surface area on the peripheral blood smear. Pathogenic variants in five erythrocyte membrane structure-related genes ANK1 (Spherocytosis, type 1; MIM#182900), SPTB (Spherocytosis, type 2; MIM#616649), SPTA1 (Spherocytosis, type 3; MIM#270970), SLC4A1 (Spherocytosis, type 4; MIM#612653) and EPB42 (Spherocytosis, type 5; MIM#612690) have been confirmed to be related to HS. There have been many studies on the pathogenic variants and mechanisms of HS, however, studies on how to manage the transmission of HS to the next-generation have not been reported. In this study, we recruited a patient with HS. Targeted next-generation sequencing with a panel of 208 genes related to blood system diseases detected a novel heterozygous variant in the SPTB: c.300+2dup in the proband. Sanger sequencing of variant alleles and haplotype linkage analysis of single nucleotide polymorphism (SNP) based on next-generation sequencing were performed simultaneously. Five embryos were identified with one heterozygous and four not carrying the SPTB variant. Single-cell amplification and whole genome sequencing showed that three embryos had varying degrees of trisomy mosaicism. One of two normal embryos was transferred to the proband. Ultimately, a healthy boy was born, confirmed by noninvasive prenatal testing for monogenic conditions (NIPT-M) to be disease-free. This confirmed our successful application of PGT in preventing transmission of the pathogenic variant allele in the HS family.

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Section: Discussionmentioning

confidence: 99%

Integrative preimplantation genetic testing analysis for a Chinese family with hereditary spherocytosis caused by a novel splicing variant of SPTB

Tian,

Wang,

Yang

et al. 2023

Front. Genet.

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“…Biopsied trophectoderm were washed thoroughly in phosphate-buffered saline (Cell Signaling Technology, Theera Trading Co. Ltd. Bangkok, Thailand) with 0.1% polyvinyl alcohol (Sigma-Aldrich, Chiangmai VM Co., Ltd., Chiang Mai, Thailand) before transferring to microcentrifuge tubes with a total volume of 4 µL. DNA extraction was performed using alkaline lysis buffer protocol [26]. Three µL of denaturation buffer (0.25 µL of 1 M DTT and 2.75 µL of buffer DLB (REPLI-g® Single Cell Kit, Chiangmai VM Co., Ltd., Chiang Mai, Thailand)) was added, and mixtures were incubated at 65 • C for ten minutes.…”

Section: Cell Lysis and Whole Genome Amplificationmentioning

confidence: 99%

“…Amplified MDA samples were tested with SNP array using Illumina HumanKaryomap-12 DNA Analysis Kit (Bio-Active Co. Ltd., Bangkok, Thailand) according to the manufacturers' instruction [22,26]. SNP genotyping information was analyzed using BlueFuse Multi software version 4.5 (Illumina, Inc. California, USA) for karyomapping analysis and molecular cytogenetics.…”

Section: Snp Array and Karyomapping Analysismentioning

confidence: 99%

“…TYR 819+3insATATGCC, TYR c.896G>A and HUMTH01 fragments were labeled with 6'FAM ® (blue), HEX ® (green), and NED ® (yellow/black) fluorescent dyes, respectively. Mini-sequencing [26] was carried out for mutation analysis of TYR c.896G>A mutation. Details of the primers are summarized in Table 1.…”

Section: Multiplex Fluorescent Pcrmentioning

confidence: 99%

“…Data were analyzed by GeneMapper ® software version 4.0. The color of individual peaks was interpreted as A (Green, dR6G dye), C (Yellow/Black, dTAMRA™ dye), G (Blue, dR110 dye), and T (Red, dROX™ dye) [26].…”

Section: Mini-sequencingmentioning

confidence: 99%

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Pre-Implantation Genetic Testing for Oculocutaneous Albinism Type 1 Using Karyomapping

Piyamongkol¹,

Mongkolchaipak²,

Chaidaroon³

et al. 2022

Clin. Exp. Obstet. Gynecol.

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Background: Oculocutaneous albinism type IA (OCA1) is the most severe form of albinism, an autosomal recessive inherited deficit of the pigment melanin causing distinctive alterations of skin, hair, and visual system. Pre-implantation genetic testing (PGT) is a substitution for prenatal diagnosis. Methods: This study accomplished SNP array with karyomapping for PGT of OCA1 and validated the results with PCR-based PGT. Results: One family with a risk of having OCA1 c.819+3insATATGCC and c.896G>A (p.R299H) offspring chose to go through karyomapping PGT. Novel PCR protocols employing fluorescent PCR and mini-sequencing were developed, tested, and applied. In the clinical PGT cycle, two blastocyst stage embryos were subjected to PGT. Karyotyping PGT results of OCA1 revealed both of the embryos to be normal. PCR analysis confirmed haplotyping results. However, copy number variation (CNV) analysis exhibited an additional chromosome 14 and segmental loss of 7q in embryo No. 1, i.e., 47, XY,+14,-7q, and an additional chromosome 22 in embryo No. 2, i.e., 47, XY,+22. Therefore, there was no appropriate embryo for transfer. The patient will return for the next PGT cycle. Conclusions: Karyomapping PGT for OCA1, including insertion c.819+3insATATGCC and point mutation c.896G>A (p.R299H), was performed alongside PCR techniques. Karyomapping gives benefits of CNV information to avoid the transfer of chromosomally unbalanced embryos.

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The successful strategy of comprehensive pre-implantation genetic testing for beta-thalassaemia–haemoglobin E disease and chromosome balance using karyomapping

Cited by 2 publications

References 21 publications

Integrative preimplantation genetic testing analysis for a Chinese family with hereditary spherocytosis caused by a novel splicing variant of SPTB

Integrative preimplantation genetic testing analysis for a Chinese family with hereditary spherocytosis caused by a novel splicing variant of SPTB

Pre-Implantation Genetic Testing for Oculocutaneous Albinism Type 1 Using Karyomapping

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