Primer combination, kit and method for detecting autosomal copy number variation

Abstract

The invention discloses a primer combination, a kit and a method for detecting autosomal copy number variation. The invention firstly provides the primer combination for detecting autosomal copy number variation. The primer combination comprises a Barcode primer, an upstream primer, a downstream outer primer and a downstream inner primer. The invention further discloses a method for detecting autosomal copy number variation. Retrotransposon regions are used as amplification specific target regions, the regions can be covered by designing one to several limited pairs of primers to enrich the whole genome, the copy number level of each region of different chromosomes can be truly reflected, a amplicon library is constructed by combining a one-step method, sample pollution is avoided, the method for detecting the autosomal copy number variation has the characteristics of simple operation, high sensitivity, high accuracy and low cost, and the application of the chromosome copy number detection method in actual detection can be truly realized.

Description

technical field [0001] The present invention relates to the field of biotechnology. Specifically relates to a primer combination, kit and method for detecting autosomal copy number variation. Background technique [0002] Studies have shown that the occurrence of tumors is related to the activation of proto-oncogenes and the inactivation of tumor suppressor genes, and chromosome copy number variation (CNV) also plays an important role in the occurrence and development of tumors. Chromosomal amplification or deletion may increase the copy number of some proto-oncogenes or decrease the copy number of tumor suppressor genes, affecting the expression of related genes. Chromosomal copy number variation exists in many tumors and is an important factor for cell carcinogenesis. index. [0003] There are many existing CNV detection technologies, such as fluorescence in situ hybridization technology, microarray technology, SNP typing chip, oligonucleotide microarray analysis technol...

AI Technical Summary

Problems solved by technology

[0003] There are many existing CNV detection technologies, such as fluorescence in situ hybridization technology, microarray technology, SNP typing chip, oligonucleotide microarray analysis technology, multiple ligation probe amplification technology, and next-generation high-throughput sequencing-based The whole genome sequencing technology of the platform, etc., although the detection technology is diverse, due to the limited detection area, cumbersome operation process, poor stability, low sensitivity, or high detection cost, etc., all limit the detection of chromosome copy number variation. application promotion

Among them: Whole-genome sequencing based on next-generation sequencing is the most effective and accurate technology for detecting chromosome copy number variation, which can truly reflect the copy number levels of different regions of chromosomes. However, the biggest problem with this technology is that the cost of detection is too high , but can not be widely used in practical applications; single nucleotide polymorphism typing chip, which analyzes the difference in copy number by comparing the average value of the hybridization signal intensity of the DNA to be tested and the hybridization signal of the control microarray, its shortcomings The SNP distribution in the high-density chip is uneven, the density of SNP probes in repetitive regions and complex regions is relatively poor, the definition is not enough, and the detection effect is poor. At the same time, the cost of this detection method is high; the multiple connection probe amplification technology is Two adjacent probes are designed according to the target site, and the 5' ends of all probes are connected with universal primers. The copy number detection is realized by comparing the difference between the test sample and the control sample. This method can quickly and sensitively realize the copy number However, this method can only detect sites with known sequences, and due to the high specificity requirements of probes, it is not suitable for the detection of all sites; fluorescence in situ hybridization technology is a classic cytological detection technology, This detection technology also designs probes for known specific areas, which can locate and accurately quantify the target area, but the detection throughput of this method is low, the cost is high, and the detection personnel are required to have rich experience; while for other detection technologies, For example, RT-qPCR compares the target gene and the reference gene for relative quantification and detects the relative copy number abundance. However, due to the low throughput and limited detection area, it is difficult to be practically applied in the chromosome copy number variation

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