Evaluation of preimplantation genetic testing for structural rearrangements based on next generation sequencing
Objective To evaluate the performance of preimplantation genetic testing(PGT)kit for structural rearrangements based on next generation sequencing using pedigrees.Methods First,we con-ducted single-cell whole-genome amplification of simulated embryonic cell samples.Next,we constructed the li-brary by performing enzyme digestion,splicing,and magnetic bead enrichment with DNA from samples in the pedigrees.We quantified the library and used a sequencer for sequencing.We then used bioinformatics software to align the acquired sequence with the human genome reference sequence and conducted SNP analysis.Based on the genetic relationship of the pedigrees,we screened effective SNP loci and constructed a haplotype map of the pedigrees.Finally,we analyzed the regions closely linked upstream and downstream of the translocation break-point to identify embryos carrying the translocation and normal embryos.Results According to the sequencing results for copy number variation(CNV)of the reference sample,the breakpoint position of chromosome 1 was chr1:194390001.The effective typing region of the embryonic sample was the upstream 1 Mb region(chr1:193390001-194390001)with 63 effective sites.The breakpoint position of chromosome 11 was chr11:13378000.The effective typing region of the embryonic sample was the upstream 1 Mb region(chr11:132780001-133780001)with 36 effective sites.The embryo sample was classified as a carrier type through haplotype clas-sification.Conclusion The evaluated preimplantation chromosomal structural abnormality detection kit can accurately detect chromosomal structural abnormalities in family samples,offering a new method of detection.
NETL
NSTL
万方数据
