Construction of a DNA molecular tag system for the safety assurance of genetic testing of preimplantation embryos
Objective:To explore the feasibility of using DNA molecular tags in the safety assurance of genetic testing of preimplantation embryos(PGT).Methods:Four types of samples for 12 cases in each type,including tripronuclear(3PN)embryos,waste cleavage-stage embryos,biopsy samples from 4CC blastocyst,and genomic DNA amplification products of actual biopsy cells,were selected form the patients who underwent IVF assisted pregnancy at the Reproductive Medicine Center of Fujian Maternal and Child Health Hospital between January and February 2024.Specific DNA molecular tags were preadded to sample collection tubes or the amplification products before amplification,copy number variation(CNV)library construction and next-generation sequencing(NGS).The DNA molecular tag sequences in the intermediate products of the whole process and the DNA molecular tag sequences in the NGS data were analyzed.Results:The Sanger sequencing results of PCR amplification products showed that the respective unique DNA molecular tag sequences were identified in the various products of all samples,and the sequences were all consistent with the added DNA molecular tag,fully consistent with the external tag.The concentrations of the amplification products and sequencing library of each sample were within the normal range.The measured DNA molecular tag sequences accounted for only approximately 1％of all NGS sequencing data,and quality control indicators of the sequencing data,including Q30,were also within their normal ranges.The embryonic genetic information obtained through conventional bioinformatics analysis is completely consistent with the results of previous sequencing analysis.Conclusions:DNA molecular tags can accurately and effectively associate with frozen biopsy embryos and the genetic test results of biopsy cells and can also track and proofread the genetic testing of biopsy cells in the whole process,thus strongly ensuring the safety of genetic testing of preimplantation embryos.
万方数据
维普
