Comparative analysis of the reliability of data on the molecular karyotype of the embryo obtained by whole-genome amplification methods based on NGS technology of trophectoderm of individual cells
- Authors: Glotov O.S.1,2,3, Saifitdinova A.F.4,5, Pavlova O.A.5,6, Leonteva O.A.5, Poliakova I.V.2, Maslennikov A.B.7
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Affiliations:
- Pediatric Research and Clinical Center for Infectious Diseases
- Serbalab LLC
- D.O. Ott Research Institute of Obstetrics, Gynecology, and Reproductology
- Herzen State Pedagogical University of Russia
- International Centre for Reproductive Medicine
- Beagle Ltd.
- Novosibirsk City Clinical Hospital
- Issue: No 6 (2024)
- Pages: 66-74
- Section: Original Articles
- URL: https://journals.eco-vector.com/0300-9092/article/view/634342
- DOI: https://doi.org/10.18565/aig.2024.78
- ID: 634342
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Abstract
Aneuploidies in human embryos play a significant role in the development of implantation failures and early reproductive losses in assisted reproductive technology (ART) programs. Preimplantation genetic testing for aneuploidy (PGT-A) is the most commonly used method for assessing embryo ploidy. The introduction of PGT-A into clinical practice has greatly reduced the chances of transferring aneuploid embryos to the uterus and has increased the effectiveness of ART programs.
Different PGT-A methods and sampling techniques have advantages and disadvantages, and there are various reasons for potential false-positive and false-negative PGT-A results. Since only a few cells are used for the study, the uniformity of DNA amplification is crucial for obtaining reliable analysis results. Therefore, the most important stage of PGT-A is whole-genome amplification (WGA).
This article presents an analysis of the current literature regarding the effectiveness and reliability of methods for amplifying small quantities of DNA. The results of our study, which compares different methods of whole-genome amplification of individual trophectoderm cells, are presented in order to obtain reliable data on the molecular karyotype of embryos using NGS. The effectiveness of using domestic WGA kits has been demonstrated.
Conclusion: Comparing PGT-A results using various kits has shown that the "hybrid" protocol we developed, which allows for the use of both foreign and domestic WGA kits, is applicable. This approach has demonstrated the high reliability of WGA kits with SD polymerase for preparing materials for PGT of human embryos, and can also be applied in other areas of biomedicine and forensic research where high reliability and accuracy in amplifying extremely small quantities of DNA is required.
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About the authors
Oleg S. Glotov
Pediatric Research and Clinical Center for Infectious Diseases; Serbalab LLC; D.O. Ott Research Institute of Obstetrics, Gynecology, and Reproductology
Author for correspondence.
Email: olglotov@mail.ru
ORCID iD: 0000-0002-0091-2224
Dr. Bio. Sci., Head of the Department of Experimental Medical Virology, Molecular Genetics and Biobanking; Senior Researcher at the Department of Genomic Medicine
Russian Federation, St. Petersburg; St. Petersburg; St. PetersburgAlsu F. Saifitdinova
Herzen State Pedagogical University of Russia; International Centre for Reproductive Medicine
Email: saifitdinova@mail.ru
ORCID iD: 0000-0002-1221-479X
Dr. Bio. Sci., Professor at the Department of Human and Animal Anatomy and Physiology; Deputy Head of the Laboratory of Assisted Reproductive Technologies
Russian Federation, St. Petersburg; St. PetersburgOlga A. Pavlova
International Centre for Reproductive Medicine; Beagle Ltd.
Email: pavlova@biobeagle.com
ORCID iD: 0000-0001-9488-6903
PhD, Biologist, Laboratory of Assisted Reproductive Technologies; Leading Specialist
Russian Federation, St. Petersburg; St. PetersburgOlga A. Leonteva
International Centre for Reproductive Medicine
Email: olga_leont@mail.ru
ORCID iD: 0000-0002-3667-0511
Embryologist, Laboratory of Assisted Reproductive Technologies
Russian Federation, St. PetersburgIrena V. Poliakova
Serbalab LLC
Email: irena.88@inbox.ru
ORCID iD: 0000-0002-5738-8443
Biologist
Russian Federation, St. PetersburgArkadiy B. Maslennikov
Novosibirsk City Clinical Hospital
Email: mab2000@mail.ru
ORCID iD: 0009-0002-8046-3816
PhD (Med), Head of the Novosibirsk Regional DNA-Diagnostics Laboratory
Russian Federation, NovosibirskReferences
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