Please use this identifier to cite or link to this item: 10.1007/s10815-018-1187-4
Title: Performance comparison of two whole genome amplification techniques in frame of multifactor preimplantation genetic testing
Authors: Volozonoka, Ludmila
Perminov, Dmitry
Korņejeva, Liene
Alkšere, Baiba
Novikova, Natālija
Pīmane, Evija Jokste
Blumberga, Arita
Kempa, Inga
Miskova, Anna
Gailīte, Linda
Fodina, Violeta
Scientific Laboratory of Molecular Genetics
Department of Obstetrics and Gynaecology
Keywords: Aneuploidy;Embryo;Preimplantation genetic testing;Single gene disorder;Whole genome amplification;3.2 Clinical medicine;1.1. Scientific article indexed in Web of Science and/or Scopus database;Reproductive Medicine;Genetics;Obstetrics and Gynaecology;Developmental Biology;Genetics(clinical)
Issue Date: 1-Aug-2018
Citation: Volozonoka , L , Perminov , D , Korņejeva , L , Alkšere , B , Novikova , N , Pīmane , E J , Blumberga , A , Kempa , I , Miskova , A , Gailīte , L & Fodina , V 2018 , ' Performance comparison of two whole genome amplification techniques in frame of multifactor preimplantation genetic testing ' , Journal of Assisted Reproduction and Genetics , vol. 35 , no. 8 , pp. 1457-1472 . https://doi.org/10.1007/s10815-018-1187-4
Abstract: Purpose: To compare multiple displacement amplification and OmniPlex whole genome amplification technique performance during array comparative genome hybridization (aCGH), Sanger sequencing, SNaPshot and fragment size analysis downstream applications in frame of multifactor embryo preimplantation genetic testing. Methods: Preclinical workup included linked short tandem repeat (STR) marker selection and primer design for loci of interest. It was followed by a family haplotyping, after which an in vitro fertilization preimplantation genetic testing (IVF-PGT) cycle was carried out. A total of 62 embryos were retrieved from nine couples with a confirmed single gene disorder being transmitted in their family with various inheritance traits—autosomal dominant (genes—ACTA2, HTT, KRT14), autosomal recessive (genes—ALOX12B, TPP1, GLB1) and X-linked (genes—MTM1, DMD). Whole genome amplification (WGA) for the day 5 embryo trophectoderm single biopsies was carried out by multiple displacement amplification (MDA) or polymerase chain reaction (PCR)-based technology OmniPlex and was used for direct (Sanger sequencing, fragment size analysis, SNaPshot) and indirect mutation assessment (STR marker haplotyping), and embryo aneuploidy testing by array comparative genome hybridization (aCGH). Results: Family haplotyping revealed informative/semi-informative microsatellite markers for all clinical cases for all types of inheritance. Indirect testing gave a persuasive conclusion for all embryos assessed, which was confirmed through direct testing. The overall allele dropout (ADO) rate was higher for PCR-based WGA, and MDA shows a better genomic recovery scale. Five euploid embryos were subjected to elective single embryo transfer (eSET), which resulted in four clinical pregnancies and birth of two healthy children, which proved free of disease causative variants running in the family postnataly. Conclusions: A developed multifactor PGT protocol can be adapted and applied to virtually any genetic condition and is capable of improving single gene disorder preimplantation genetic testing in a patient-tailored manner thus increasing pregnancy rates, saving costs and increasing patient reliability.
Description: Publisher Copyright: © 2018, The Author(s).
DOI: 10.1007/s10815-018-1187-4
ISSN: 1058-0468
Appears in Collections:Research outputs from Pure / Zinātniskās darbības rezultāti no ZDIS Pure

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