Detection of DNA in Human Blastocyst Cavity Aspirate by Multiplex PCR
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Preimplantation genetic testing (PGT) is a modern method of detection of chromosomal and genetic abnormalities in a human embryo before its transfer to the uterus. The genetic material is obtained by embryo biopsy. Here, we attempted to evaluate the efficiency of a method of noninvasive biopsy—aspiration of the blastocoel contents (blastocentesis) of human embryos. In this study, a biopsy was carried out human embryos with low morphological characteristics (3–4 CC according to Gardner grading) on day 6–7 of development. DNA obtained from the aspirate, as well as from the blastocyst, was analyzed by QF-PCR (quantitative fluorescent polymerase chain reaction) after whole genome amplification. In total, 24 blastocysts and aspirate samples obtained from them were analyzed; assayable DNA was found in seven (29%) aspirate samples from the blastocyst cavity, and this DNA was identical to blastocyst DNA in five (71%) cases. Thus, it was shown that, using aspiration of the blastocoel fluid of the human embryo, it is possible to obtain DNA suitable for analysis by molecular genetic methods. The features and advantages of the use of multiplex QF-PCR method combined with whole genome amplification for studying DNA obtained during aspiration of the blastocoel fluid are discussed. The prospects of DNA obtainment by the noninvasive biopsy method for preimplantation genetic testing (PGT) in the routine practice of infertility treatment and prevention of chromosomal and genetic abnormalities in newborns are considered.
Keywordshuman embryo PGT embryo biopsy blastocyst cavity blastocentesis
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- Bazanov, P.A., Mityushina, N.G., Yutkin, E.V., et al., Optimization of the management of patients when performing preimplantation genetic diagnosis, Vestn. Ross. Univ. Druzhby Narodov, Ser.: Med., 2013, no. 5, pp. 97–102.Google Scholar
- Kumar, G., Garnova, E., Reagin, M., et al., A improved multiple displacement amplification with phi29 DNA polymerase for genotyping of single human cells, Bio-Techniques, 2008, vol. 44, no. 7, pp. 879–890.Google Scholar