Detection reagent kit for methylation of septin 9 genes in human peripheral blood circulation tumor DNA

Abstract

The invention discloses a detection reagent kit for methylation of septin 9 genes in human peripheral blood circulation tumor DNA. The detection reagent kit comprises methylation-specific primers for target loci of the septin 9 genes, non-methylation-specific dideoxycytidine modified primers for the target loci of the septin 9 genes, hydrolysis probes specific to the septin 9 genes, primers for reference genes and hydrolysis probes for the reference genes, wherein the target loci of the septin 9 genes are at least one CpG locus contained in an exon region of transcripts gamma 1 of the septin 9 genes V2, and the reference genes are one or more of GAPDH, beta-actin, B2M, SDHA and HPRT1. By means of the component configuration scheme and the result interpretation scheme of the reagent kit, the detection process is easy and convenient, and operability is high. Detection results are reliable, and specificity is good.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a kit for detecting methylation of septin9 gene in human peripheral blood circulating tumor DNA. Background technique [0002] Colorectal cancer is one of the major diseases that cause human death worldwide. In Europe and the United States, colorectal cancer ranks second and third, respectively. In 2012, there were about 447,000 new cases of colorectal cancer in Europe, and about 215,000 deaths. In 2013, the number of new cases in the United States was about 142,820 people, and about 50,830 deaths. In China, the number of new cases of colorectal cancer in 2012 was about 400,000, and it has become the disease with the third highest morbidity and mortality among Chinese people. [0003] Early detection of colorectal cancer can be achieved through routine screening, and early treatment can be carried out to prevent the progression of the disease or even cure it. However, t...

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Problems solved by technology

In China, the screening rates of fecal occult blood test and colonoscopy are estimated to be less than 5% and 3% respectively

This is a classic methylation research method. Its advantages are: relatively simple, low cost, clear methylation sites, and easy interpretation of experimental results. The disadvantages are: 1. Since CG is not limited to CCGG sequences, it is not The CG in this sequence will be ignored; 2. Only when the methylation status of key sites related to transcription is detected, the results of this detection method are meaningful; 3. Relatively speaking, the Southern method is more complicated and requires The amount of sample is large; 4. There is a problem of false positives caused by incomplete enzyme digestion; 5. It is not suitable for mixed samples

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