Method for simultaneously completing gene locus, chromosome and linkage analysis

Abstract

The invention relates to a method for simultaneously completing gene locus, chromosome and linkage analysis. The method concretely and mainly comprises the following steps: collecting an embryo cell sample, amplifying a whole genome, amplifying a target gene mutation locus, establishing a whole genome and target gene mutation locus library, carrying out high flux sequencing, and carrying out data analysis. Multiple-item comprehensive detection is completed through one step by combining a whole genome amplification technology with the high flux sequencing, so respective detection of single-gene genetic disease mutation site, chromosome diseases and linkage analysis through using multiple methods and multiple steps is avoided. The method provided by the invention provides favorable conditions for a tiny amount of a sample, can be used for PGD detection to determine whether an embryo carries a pathogenic gene and chromosome copy number abnormity or not, is also suitable for genetic screening of embryos of recurrent abortion older women, and realizes multi-item detection of a plurality of single samples through one step. The method has the advantages of simple operation, short period and strong feasibility, so promotion and application of the method are facilitated.

Description

technical field [0001] The invention relates to the fields of genome sequence analysis and bioinformatics, and specifically relates to a method for completing single-cell single-gene disease, chromosomal disease and linkage analysis in one step by using MALBAC amplification technology combined with high-throughput sequencing. Background technique [0002] Genetic diseases refer to diseases caused by changes in the genetic material (gene sequences on chromosomes or mitochondrial DNA) in the human body. In recent years, the incidence of genetic diseases has increased year by year. In my country, as many as tens of thousands of children with chromosomal abnormalities are born every year, and children with chromosomal abnormalities are still incurable. At present, there are more than 7,000 kinds of single-gene genetic diseases that have been discovered, and more than 4,000 of which have been identified as pathogenic genes. The overall morbidity rate in infants and the overall p...

AI Technical Summary

Problems solved by technology

Multiplex fluorescent PCR technology combines multiplex PCR with fluorescent probes to detect STR typing. Due to the lack of STR linkage markers, there may be no available STR sites in specific cases. Therefore, a lot of pre-experimental work is required before clinical testing. STR genetics Most of the markers are far away from the disease-causing site, prone to chromosomal recombination and misdiagnosis, and the detection cost is high, so this technology cannot be used for batch detection

Compared with the method of capturing SNP by chips and probes, the cost is relatively high and the cycle is relatively long compared with the method of capturing SNP by ordinary PCR

[0005] Existing technology cannot simultaneously complete the three items of monogenic genetic disease, chromosomal abnormality and linkage analysis through one-step detection, and the whole genome sample obtained from a small number of cells is prone to allelic dropout (ADO) when multiple tests are performed separately during the experiment , pollution and other phenomena, this field is in urgent need of a technical solution that can solve this problem

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