scholarly journals PREIMPLANTATION GENETIC TESTING: Clinical experience of preimplantation genetic testing

Reproduction ◽  
2020 ◽  
Vol 160 (5) ◽  
pp. A45-A58
Author(s):  
Martine De Rycke ◽  
Veerle Berckmoes ◽  
Anick De Vos ◽  
Stefanie Van De Voorde ◽  
Pieter Verdyck ◽  
...  
Keyword(s):  
Genetic Testing ◽  
Whole Genome Amplification ◽  
Whole Genome ◽  
Preimplantation Genetic Testing ◽  
Hormonal Stimulation ◽  
Testing Tools ◽  
Technological Advances ◽  
Genome Wide ◽  
Wide Testing

Thirty years of rapid technological advances in the field of genetic testing and assisted reproduction have reshaped the procedure of preimplantation genetic testing (PGT). The development of whole genome amplification and genome-wide testing tools together with the implementation of optimal hormonal stimulation protocols and more efficient cryopreservation methods have led to more accurate diagnoses and improved clinical outcomes. In addition, the shift towards embryo biopsy at day 5/6 has changed the timeline of a typical PGT clinical procedure. In this paper, we present an up-to-date overview of the different steps in PGT from patient referral to baby follow-up.

scholarly journals Performance comparison of two whole genome amplification techniques in frame of multifactor preimplantation genetic testing

2018 ◽  
Vol 35 (8) ◽  
pp. 1457-1472 ◽  
Author(s):  
Ludmila Volozonoka ◽  
Dmitry Perminov ◽  
Liene Korņejeva ◽  
Baiba Alkšere ◽  
Natālija Novikova ◽  
...  
Keyword(s):  
Genetic Testing ◽  
Whole Genome Amplification ◽  
Performance Comparison ◽  
Whole Genome ◽  
Preimplantation Genetic Testing ◽  
Genome Amplification

Single cell and whole genome amplification product validation of preimplantation genetic testing systems for monogenic disorders

2020 ◽  
pp. 63-64
Author(s):  
Е.В. Соловьёва ◽  
О.Р. Канбекова ◽  
Д.И. Жигалина ◽  
Н.А. Скрябин ◽  
Л.И. Минайчева
Keyword(s):  
Genetic Testing ◽  
Single Cell ◽  
Whole Genome Amplification ◽  
Single Cells ◽  
Whole Genome ◽  
Amplification Product ◽  
Preimplantation Genetic Testing ◽  
Monogenic Disorders ◽  
Pathogenic Variants ◽  
Whole Genome Amplification Product

В рамках подготовительных этапов преимплантационного тестирования 9 моногенных заболеваний (ПГT-М) методом гнездовой ПЦР проанализированы 144 локуса (STR и патогенные варианты)109 единичных клеток и 24 образцов продуктов полногеномной амплификации (ПГА) нескольких клеток. Pre-examination single cell validation was performed for preimplantation genetic testing (PGT-M) for 9 monogenic disorders by nested PCR for STR and pathogenic variants. Totally 109 single cells and 24 WGA products (by MDA) were analyzed.

scholarly journals Whole-genome amplification/preimplantation genetic testing for propionic acidemia of successful pregnancy in an obligate carrier Mexican couple: A case report

2021 ◽  
Vol 9 (29) ◽  
pp. 8797-8803
Author(s):  
Adina Neumann ◽  
Miguel Angel Alcantara-Ortigoza ◽  
Ariadna González-del Angel ◽  
Nestor Alejandro Zarate Díaz ◽  
Javier Sam Santana ◽  
...  
Keyword(s):  
Case Report ◽  
Genetic Testing ◽  
Whole Genome Amplification ◽  
Propionic Acidemia ◽  
Whole Genome ◽  
Obligate Carrier ◽  
Successful Pregnancy ◽  
Preimplantation Genetic Testing ◽  
Genome Amplification

P–540 A feasible diagnostic approach for the cryptic subtelomeric traslocations in early recurrent miscarriage patients by preimplantation genetic testing (PGT)

2021 ◽  
Vol 36 (Supplement_1) ◽  
Author(s):  
B Lledo ◽  
R Morales ◽  
J A Ortiz ◽  
A Cascales ◽  
A Fabregat ◽  
...  
Keyword(s):  
Genetic Testing ◽  
Recurrent Miscarriage ◽  
Chromosome Translocation ◽  
Accurate Diagnosis ◽  
Molecular Techniques ◽  
Telomeric Region ◽  
Whole Genome ◽  
Preimplantation Genetic Testing ◽  
Uterine Anomalies ◽  
Cryptic Translocation

Abstract Study question Could cryptic subtelomeric traslocations in early recurrent miscarriage patients be diagnosed by preimplantation genetic testing? Summary answer PGT is a powerful tool to detect subtelomeric cryptic traslocations identifying the cause of early recurrent miscarriage and allowing subsequent genetic counselling. What is known already: Chromosome translocations are frequently associated with birth defects, spontaneous early pregnancy losses and infertility. However, submicroscopic traslocations (so-called cryptic traslocations) are too small to be detected by conventional karyotyping.. Due to balanced status, high resolution molecular techniques as arrayCGH are not able to detect it. Thus, cryptic traslocations detection is challenging. PGT is able to detect CNVs at higher resolution than routine karyotyping. Therefore, the recurrent diagnosis of CNV at embryo level could suggest a subchromosomal parental traslocation. The aim of this study is to investigate the feasibility of using PGT as an indicator of parental balanced cryptic traslocations. Study design, size, duration We included three couples who underwent PGT for unexplained repeated pregnancy loss (RPL) in our clinic from February 2020 to November 2020. Common established causes of RPL (uterine anomalies, antiphospholipid syndrome, immunological, hormonal and metabolic disorders) were previously rouled-out. Even couple karyotypes were normal. Twenty-three embryos from those couples were biopsied at blastocyst and analysed for CNVs detection using low coverage whole genome NGS. Participants/materials, setting, methods PGT by NGS was performed by Veriseq-NGS (Illumina), with previous whole genome amplification. Fluorescence in situ hybridization (FISH) using parental blood samples were performed to validate the origin of subchromosomal number variation. Commercially available subtelomeric specific probes were selected according to the CNV identified and the procedures were performed according to the manufacturer’s protocols. Main results and the role of chance Overall, CNVs of terminal duplication and deletion that imply unbalanced traslocation derivatives were detected in the 43.5% of biopsied embryos. For couple 1, 4 out of 5 embryos (80%) carried deletion of telomeric region on chromosomes 5 and 21. Three out of 6 biopsed embyos (50%) were diagnosed with subchromosomal copy variants at telomeric region on chromosomes 6 and 16 for couple 2. In the case of couple 3, three out of 12 embryos (25%) were carriers of CNV at subtelomeric region on chromosomes 2 and 6. The size of CNVs detected ranges from 8Mb to 20Mb. Accurate diagnosis with the parental study was made by FISH. The combination of probes to detect the structural chromosome alteration were: Tel5qter-LSI21q, Tel6pter-CEP16 and Tel6pter-CEP6 for each couple respectively. The FISH studies reveal that CNVs were inherited from one parent carrying the balanced cryptic traslocation. Ultimately, the abnormal karyotype from the carrier parent were 46,XY,t(5;21)(q33.2;q21.2) for couple 1, 46,XY,t(6;16)(p22.3;q22.1) for couple 2 and 46,XY,t(2;6)(p25.1;p24.2) for couple 3. Finally, each couple performed a cryotransfer of a single normal balanced embryo. Two pregnancies are ongoing. Limitations, reasons for caution The main limitation of this approach is the NGS- PGT resolution. CNVs smaller than 5Mb could not be detected. Wider implications of the findings: This study shows the value of PGT for unexplained RPL, followed by parental FISH to better characterize CNVs and identify couples in whom one partner carries a cryptic translocation. Accurate diagnosis of parental chromosome translocation can achieve with FISH only, but FISH would not be performed unless PGT showed CNVs. Trial registration number Not applicable

scholarly journals DNA Analysis by Restriction Enzyme (DARE) enables concurrent genomic and epigenomic characterization of single cells

2019 ◽  
Vol 47 (19) ◽  
pp. e122-e122
Author(s):  
Ramya Viswanathan ◽  
Elsie Cheruba ◽  
Lih Feng Cheow
Keyword(s):  
Dna Methylation ◽  
Restriction Enzyme ◽  
Copy Number ◽  
Dna Analysis ◽  
Whole Genome Amplification ◽  
Single Cells ◽  
Whole Genome ◽  
Copy Number Alterations ◽  
Genome Wide

Abstract Genome-wide profiling of copy number alterations and DNA methylation in single cells could enable detailed investigation into the genomic and epigenomic heterogeneity of complex cell populations. However, current methods to do this require complex sample processing and cleanup steps, lack consistency, or are biased in their genomic representation. Here, we describe a novel single-tube enzymatic method, DNA Analysis by Restriction Enzyme (DARE), to perform deterministic whole genome amplification while preserving DNA methylation information. This method was evaluated on low amounts of DNA and single cells, and provides accurate copy number aberration calling and representative DNA methylation measurement across the whole genome. Single-cell DARE is an attractive and scalable approach for concurrent genomic and epigenomic characterization of cells in a heterogeneous population.

scholarly journals Preimplantation Genetic Testing for Monogenic Disorders

Genes ◽  
2020 ◽  
Vol 11 (8) ◽  
pp. 871 ◽  
Author(s):  
Martine De Rycke ◽  
Veerle Berckmoes
Keyword(s):  
Genetic Testing ◽  
Whole Genome Amplification ◽  
State Of The Art ◽  
Snp Array ◽  
Genetic Data ◽  
Monogenic Disorder ◽  
Preimplantation Genetic Testing ◽  
Monogenic Disorders ◽  
The Cost ◽  
Concurrent Analysis

Preimplantation genetic testing (PGT) has evolved into a well-established alternative to invasive prenatal diagnosis, even though genetic testing of single or few cells is quite challenging. PGT-M is in theory available for any monogenic disorder for which the disease-causing locus has been unequivocally identified. In practice, the list of indications for which PGT is allowed may vary substantially from country to country, depending on PGT regulation. Technically, the switch from multiplex PCR to robust generic workflows with whole genome amplification followed by SNP array or NGS represents a major improvement of the last decade: the waiting time for the couples has been substantially reduced since the customized preclinical workup can be omitted and the workload for the laboratories has decreased. Another evolution is that the generic methods now allow for concurrent analysis of PGT-M and PGT-A. As innovative algorithms are being developed and the cost of sequencing continues to decline, the field of PGT moves forward to a sequencing-based, all-in-one solution for PGT-M, PGT-SR, and PGT-A. This will generate a vast amount of complex genetic data entailing new challenges for genetic counseling. In this review, we summarize the state-of-the-art for PGT-M and reflect on its future.

scholarly journals A whole genome RNAi screen of Drosophila S2 cell spreading performed using automated computational image analysis

2010 ◽  
Vol 191 (3) ◽  
pp. 471-478 ◽  
Author(s):  
Michael V. D'Ambrosio ◽  
Ronald D. Vale
Keyword(s):  
Image Data ◽  
Cell Spreading ◽  
Rnai Screen ◽  
Whole Genome ◽  
Casein Kinase I ◽  
Drosophila Melanogaster S2 Cells ◽  
Technological Advances ◽  
Genome Wide ◽  
Computational Image Analysis ◽  
Rate Limiting

Recent technological advances in microscopy have enabled cell-based whole genome screens, but the analysis of the vast amount of image data generated by such screens usually proves to be rate limiting. In this study, we performed a whole genome RNA interference (RNAi) screen to uncover genes that affect spreading of Drosophila melanogaster S2 cells using several computational methods for analyzing the image data in an automated manner. Expected genes in the Scar-Arp2/3 actin nucleation pathway were identified as well as casein kinase I, which had a similar morphological RNAi signature. A distinct nonspreading morphological phenotype was identified for genes involved in membrane secretion or synthesis. In this group, we identified a new secretory peptide and investigated the functions of two poorly characterized endoplasmic reticulum proteins that have roles in secretion. Thus, this genome-wide screen succeeded in identifying known and unexpected proteins that are important for cell spreading, and the computational tools developed in this study should prove useful for other types of automated whole genome screens.

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