Primer probe system for SEPT9 (septin-9) gene promoter methylation detection and kit adopting primer probe system

Abstract

The invention relates to a primer probe system for SEPT9 (septin-9) gene promoter methylation detection and a kit adopting the primer probe system. The primer probe system comprises a primer probe set X used for determining genomic DNA quality, a primer probe set Y used for determining a methylation conversion rate and a primer probe set Z used for detecting SEPT9 gene promoter methylation conditions, wherein the primer probe set X comprises a forward primer a, a reverse primer a and a probe a; the primer probe set Y comprises a forward primer b, a reverse primer b and a probe b; the primer probe set Z comprises the forward primer b, the reverse primer b and a probe c; fluorescence report groups are arranged at 5' ends of the probe a, the probe b and the probe c, and fluorescence quenching groups are arranged at 3' ends of the probe a, the probe b, the probe c and the probe d. According to the kit, the implementing scheme is concise, and the sensitivity and the accuracy rate are high.

Description

technical field [0001] The invention relates to a gene mutation detection product, a detection primer and a detection system used in the product, and belongs to the field of biotechnology. Background technique [0002] Colorectal cancer is a malignant tumor originating from the mucosal epithelium of the large intestine; colorectal cancer can be divided into rectal cancer and colon cancer according to the location of the tumor. There are as many as 930,000 new patients with colorectal cancer every year in the world, and it has become the third most common malignant tumor in the world. With the improvement of people's living standards and changes in eating habits, the incidence of colorectal cancer in China is spiraling at a rate of 4.2%, far exceeding the international level of 2%. In my country, there are 130,000 to 160,000 new cases every year, and the incidence of colorectal cancer in digestive tract tumors is second only to gastric cancer; in my country, the prevalence o...

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Problems solved by technology

The MSP method combined with the nested PCR amplification method can improve the specificity of the single MSP method, but the operation time is long, and compared with the MSP method, there are also false positives caused by incomplete bisulfite treatment. The pH value in the modification process is relatively absolute. Accurate, all reagents require fresh configuration, and need to repeatedly explore to find the appropriate reaction time, resulting in cumbersome detection process, and can not achieve quantitative detection, there is a high false positive rate; there is also excessive sulfite treatment, resulting in amplification 3. Methylation Sensitivity Melting Curve Analysis (MS-High Resolution Melting Curve, MS-HRM), which is cumbersome to operate, has a high false positive rate, and is not suitable for the detection of a large number of samples; 4. Fluorescence method (Methylight) , due to the limitations of the probe, there is generally no suitable control system, and the false positive rate is high; 5. Bisulfite sequencing PCR (BSP), which is a PCR combined with Sanger sequencing technology, has accurate results, but the process is cumbersome. Not suitable for high-volume testing, and too expensive to be widely used

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