Detection method and kit for detecting 1p/19q combination loss of heterozygosity of chromosome

A technology for lack of heterozygosity and detection methods, applied in the direction of microbial determination/inspection, biochemical equipment and methods, etc., can solve the problems of unreliable analysis of markers, etc., achieve fast turnaround time, high sensitivity and specificity, and easy operation easy effect

Pending Publication Date: 2019-04-23
CARRIER GENE TECH SUZHOU CO LTD +1
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Problems solved by technology

However, most next-generation sequencing platforms cannot reliably analyze these markers due to the repetitive nature of microsatellites

Method used

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  • Detection method and kit for detecting 1p/19q combination loss of heterozygosity of chromosome
  • Detection method and kit for detecting 1p/19q combination loss of heterozygosity of chromosome
  • Detection method and kit for detecting 1p/19q combination loss of heterozygosity of chromosome

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Embodiment 1

[0033] Example 1 High-throughput next-generation sequencing detection method of Chr1p / 19q co-LOH based on multiple amplicons

[0034] 1. Site selection

[0035] Select 10 SNPs on the short arm of chromosome 1 (1p) and the long arm of chromosome 19 (19q), among which, 5 SNPs on 1p are located in the 1p36.3 segment, and 5 SNPs on 19q are located in the 19q13.3 segment Segment, these two regions are used to determine whether loss of heterozygosity occurs in the common deletion segment, combined with other SNPs, it can be used to determine whether loss of heterozygosity occurs in the entire long arm or short arm. The heterozygosity distribution of each SNP in the East Asian population refers to the gnomAD_exome_EAS database. The selection criteria for SNPs are: the probability of the SNP being heterozygous is close to 50% in the population to avoid the situation that all SNPs are homozygous; the distance between SNPs in the genome is greater than 300kb to minimize the linkage bet...

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Abstract

The invention discloses a detection method for detecting 1p/19q combination loss of heterozygosity of chromosome, which comprises the following steps: 1) selecting SNP sites with the same number on 1pand 19q; 2) designing an original primer, modifying a T of each original primer, closest to the 3'end but not at the tail end, into U, if the tail end of the 3'end is T, modifying a second T towardsthe 5'direction into U to obtain a multiplex amplification primer sequence; 3) synthesizing a multiplex amplification primer and dissolving and mixing the multiplex amplification primer; 4) carrying out multiplex PCR amplification; 5) processing the amplification product to enable a single chain of U to form an AP site, and then exposing the 5'phosphoric acid tail end and 3'phosphoric acid tail end of the AP site; 6) sequencing, and judging whether 1p/19q combination loss of heterozygosity occurs on the chromosome according to SNP allelic type frequency. According to the invention, the NGS method based on multiplex amplification is used for detecting Chr1p/19qco-LOH, not only is the detection sensitivity high, and are the specificity and the efficiency high, but also the limitation on samples is small.

Description

technical field [0001] The invention relates to the field of biomedicine, in particular to the detection of chromosome 1p / 19q combined heterozygosity loss and a kit for the detection. Background technique [0002] Many important genes closely related to cell growth regulation, proliferation and differentiation are concentrated in the 1p36 at the end of the short arm of human chromosome 1 and the 19q13.3 region of the long arm of chromosome 19. In glioma, the combined loss of heterozygosity of chromosome 1p / 19q (Chr1p / 19q co-LOH) has important clinical significance. [0003] Chr1p / 19q co-LOH is associated with oligodendroglioma in histological classification. In the 2016 WHO classification of central nervous system tumors, molecular pathological detection of this indicator is recommended (see figure 1 ), this indicator is of great significance for the pathological classification of low-grade glioma, and is an important basis for the pathological classification of low-grade ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6886C12Q1/6806C12Q1/6869
CPCC12Q1/6806C12Q1/6869C12Q1/6886C12Q2600/156C12Q2531/113C12Q2537/143C12Q2521/531C12Q2521/301C12Q2535/122
Inventor 魏金旺张敖王晨戴春许强
Owner CARRIER GENE TECH SUZHOU CO LTD
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