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

Autor: Inga Kempa, Linda Gailīte, Violeta Fodina, Ludmila Volozonoka, Dmitry Perminov, Anna Miskova, Baiba Alksere, Liene Korņejeva, Natālija Novikova, Evija Jokste Pīmane, Arita Blumberga
Jazyk: angličtina
Rok vydání: 2018
Předmět:
0301 basic medicine
Male
Pregnancy Rate
Fertilization in Vitro
Biology
03 medical and health sciences
symbols.namesake
0302 clinical medicine
Pregnancy
medicine
Genetics
Single Embryo Transfer
Humans
Genetic Testing
Allele
Genetics (clinical)
Preimplantation Diagnosis
Genetic testing
Whole Genome Amplification
Sanger sequencing
Comparative Genomic Hybridization
030219 obstetrics & reproductive medicine
Preimplantation genetic testing
Single gene disorder
medicine.diagnostic_test
Tripeptidyl-Peptidase 1
Haplotype
Multiple displacement amplification
Obstetrics and Gynecology
General Medicine
Aneuploidy
Human genetics
Whole genome amplification
030104 developmental biology
Blastocyst
Reproductive Medicine
Embryo
symbols
Microsatellite
Female
Nucleic Acid Amplification Techniques
Developmental Biology
Zdroj: Journal of Assisted Reproduction and Genetics
ISSN: 1573-7330
1058-0468
Popis: 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. Electronic supplementary material The online version of this article (10.1007/s10815-018-1187-4) contains supplementary material, which is available to authorized users.
Databáze: OpenAIRE
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