Single-gene defect detection method, primer composition for single-gene defect detection in Xq28 region and kit

Abstract

The invention provides a single-gene defect detection method, a primer composition for single-gene defect detection in a Xq28 region and a kit. The method comprises the following steps: designing the primer composition; acquiring DNA (deoxyribonucleic acid) samples of a proband, parents and offspring; carrying out amplification by using the primer composition, constructing a library and carrying out sequencing analysis; and obtaining haplotypes of the proband, the parents and the offspring based on a sequencing analysis result, and determining a detection result of the single-gene defect of the offspring. The design method of the primer composition comprises the following steps: acquiring SNP loci of a to-be-detected DNA fragment, screening SNP detection loci and designing primer pairs; or obtaining the SNP loci and the STR loci of the to-be-detected DNA fragment, screening the SNP detection loci and the STR detection loci, and designing the primer pairs. The method, the primer composition and the kit provided by the invention can be used for identifying whether the gene in the to-be-detected DNA fragment of an embryo before implantation varies or not, and the qualified embryo is efficiently and accurately screened for implantation, so that the disease risk is reduced.

Description

technical field [0001] The invention relates to the technical field of gene detection, in particular to a method for detecting a single gene defect, a primer composition and a kit for detecting a single gene defect in the Xq28 region. Background technique [0002] A genetic disease is a family disease, which refers to a disease caused by a mutation (or aberration) in the genetic material (chromosomes and genes) of germ cells or fertilized eggs. Single-gene disease detection is carried out in advance, and embryos without disease-causing mutations are selected for transplantation, so as to avoid the pain caused by genetic diseases in future generations. [0003] At present, the detection of single gene diseases in preimplantation embryos requires the detection of SNP sites (single nucleotide variation sites) within about 4 Mb upstream and downstream of the target gene. For example, the Xq28 region is located on the X chromosome, which contains SLC6A8, ABCD1, L1CAM, AVPR2, MEC...

AI Technical Summary

Problems solved by technology

However, the following problems are usually encountered when using STR to construct haplotypes: the slippage phenomenon seriously affects the accuracy of STR detection and analysis; currently, STR haplotypes are mainly constructed by capillary electrophoresis, and the throughput of the detection scheme is low. There may be a problem of signal interference between them; there are few STR sites that have been published so far, and it is difficult to meet the needs of different clinical applications

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