Method for identifying balanced translocation breaking point and balanced translocation carrying state of embryo

Abstract

The invention relates to the technical field of biotechnology and relates to the field of genome sequence analysis, and particularly relates a method for identifying a balanced translocation breaking point and a balanced translocation carrying state of an embryo. The method comprises the following steps: amplifying and sequencing a sample; comparing a sequencing sequence to a reference genome and analyzing a copy number; accurately determining a position of a translocation breaking point; detecting SNP (Single Nucleotide Polymorphism) at the periphery of the breaking point; carrying out SNP genotyping; carrying out embryo haplotype analysis; comprehensively judging a regular chromosome and a translocation chromosome haplotype; determining an embryo carrying state; and selecting an embryo without balanced translocation according to a haplotype. According to the method provided by the invention, the position of the balanced translocation breaking point can be accurately determined, and a few of SNP sites can also be used for accurately judging and typing; and the embryo without the balanced translocation is identified.

Description

technical field [0001] The invention relates to the field of biotechnology, to the field of genome sequence analysis, and in particular to a method for identifying the breaking point of embryo balanced translocation and the carrying state of balanced translocation. Background technique [0002] In the scientific research and clinical application fields of biomedicine and reproductive genetics, balanced translocation is a very common abnormal chromosome structure defect in newborns, accounting for about 1 / 500-1 / 625 newborns. Balanced translocation carriers usually have a normal phenotype, and some have genetic variations such as microduplications, deletions, and gene damage, which can lead to diseases such as autism, mental retardation, and congenital deformities. Balanced translocation carriers are more likely to produce unbalanced gametes when they give birth to offspring, which can lead to recurrent miscarriage and even infertility. Therefore, in order to block the balanc...

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Problems solved by technology

[0004] However, the resolution of comparative genomic hybridization technology is relatively low, Mb level, low throughput, high cost; fluorescence in situ hybridization, only for specific positions, low resolution, unstable probe hybridization efficiency, because balanced translocation involves almost every Each zone of the chromosome, so each balanced translocation carrier needs to design a probe separately, so it will be time-consuming and costly, and cannot be used as a general detection technology; the SNP chip is designed for the whole genome, and the SNP distribution is not Will be absolutely homogeneous, so there is uncertainty about the valid sites around the balanced translocation breakpoint that can be used for linkage analysis, resulting in the inability to distinguish whether the embryo is a balanced translocation carrier

[0005] In addition to the above-mentioned technical defects, none of the above detection techniques can accurately determine the position of the breakpoint of the balanced translocation. If the position of the breakpoint is not accurate enough and exceeds a certain range, due to recombination and exchange, it will lead to the judgment of the embryo carrying / not carrying the balanced translocation. mistake

[0006] Although the technology of microdissection combined with next-generation sequencing (MicroSeq-PGD) can accurately determine the position of the balanced translocation breakpoint, it needs to go through stages such as cell culture and microdissection. The operation is complicated, the detection cycle is very long, and the price is expensive. The requirements for personnel and equipment are high, and it cannot be promoted on a large scale

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