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A reagent for early diagnosis of colorectal cancer based on combined detection of methylation levels of sdc2 and sfrp2 genes

A colorectal cancer and methylation technology, applied in the field of molecular biology, can solve the problems of time-consuming, laborious, invasive, and low accuracy, and achieve the effect of convenient and simple sampling process, reducing pollution, and reducing incidence and mortality

Active Publication Date: 2021-12-14
SHANGHAI REALBIO TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0003] In order to solve the problems of invasiveness, time-consuming and laborious, and low accuracy existing in the existing colorectal cancer diagnosis technology, the present invention provides a non-invasive early diagnosis of colorectal cancer based on the joint detection of the methylation levels of SDC2 gene and SFRP2 gene Reagents and methods to achieve accurate and simple diagnosis of early colorectal cancer

Method used

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  • A reagent for early diagnosis of colorectal cancer based on combined detection of methylation levels of sdc2 and sfrp2 genes
  • A reagent for early diagnosis of colorectal cancer based on combined detection of methylation levels of sdc2 and sfrp2 genes
  • A reagent for early diagnosis of colorectal cancer based on combined detection of methylation levels of sdc2 and sfrp2 genes

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Experimental program
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Effect test

Embodiment 1

[0085] Embodiment 1. Preparation and use of kit

[0086] In the kit of the present invention, including SDC2 gene detection primers and probes, SFRP2 gene detection primers and probes, the nucleotide sequences thereof are shown in Table 1-1 below:

[0087] Table 1-1

[0088]

[0089] The kit can also include a control, which is a β-actin detection primer and a probe, and its nucleotide sequence is shown in Table 1-2 below:

[0090] Table 1-2

[0091]

[0092] experiment procedure:

[0093] 1. DNA Extraction from Stool Samples

[0094] The specific extraction method may include the following steps: follow the standard operating procedure of the QIAamp Fast DNA Stool Mini Kit kit: add 1ml InhibitEX Buffer to the sample, vortex for 1min until the sample is fully homogenized, and centrifuge at 16000g for 1min; take 25μl proteinase K into a new Centrifuge the tube, and pipette 600μl of the supernatant from the previous step to the proteinase K tube, add 600μl Buffer AL an...

Embodiment 2

[0121] Example 2. The kit detects samples from patients with intestinal cancer

[0122] 1. DNA Extraction from Stool Samples

[0123] The stool samples of 38 patients diagnosed as colon cancer by colonoscopy and pathological examination were clinically collected, and the stool samples of 30 patients diagnosed as normal by colonoscopy were collected. DNA extraction was performed on the obtained stool samples.

[0124] The specific extraction method may include the following steps: Follow the standard operating procedure of the QIAamp Fast DNAStool Mini Kit kit: add 1ml InhibitEX Buffer to the sample, vortex for 1min until the sample is fully homogenized, and centrifuge at 16000g for 1min; take 25μl proteinase K into a new centrifuge tube, and pipette 600 μl of the supernatant from the previous step to the proteinase K tube, add 600 μl Buffer AL and vortex for 15 seconds, incubate at 70°C for 10 minutes; add 600 μl of ethanol, vortex and mix well, divide into three QIAamp spin ...

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Abstract

The invention belongs to the field of molecular biology, and in particular relates to an early diagnosis reagent for colorectal cancer based on joint detection of the methylation levels of SDC2 and SFRP2 genes. In the present invention, feces are used as detection samples, and the combination of SDC2 gene and SFRP2 gene is used as a biomarker to detect the methylation levels of SDC2 gene and SFRP2 gene. The results showed that the methylation levels of SDC2 and SFRP2 genes detected in feces maintained a high degree of correlation with the incidence of colorectal cancer. The detection sensitivity of colorectal cancer was 94.74%, and the specificity was 96.67%. The detection method of the invention is accurate, fast and simple, and can realize non-invasive colorectal cancer screening and early diagnosis.

Description

technical field [0001] The invention belongs to the field of molecular biology, and in particular relates to an early diagnosis reagent for colorectal cancer based on joint detection of the methylation levels of SDC2 and SFRP2 genes. Background technique [0002] Colorectal cancer is one of the most common malignant tumors in the world. Early screening and diagnosis of colorectal cancer in the population is the key to disease prevention and treatment. Currently, the clinical screening of colorectal cancer is mainly carried out through fecal occult blood test and colonoscopy. Fecal occult blood test is to detect the hemoglobin in the stool. If multiple and continuous positive reactions indicate gastrointestinal bleeding, further examination should be done to alert the occurrence of intestinal tumors. Although this technique is convenient and non-invasive, it is easily affected by diet and other factors, which in turn affects the accuracy of the test results. In addition, co...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6886
CPCC12Q1/6886C12Q2600/154
Inventor 王美丛刘晶秦楠
Owner SHANGHAI REALBIO TECH CO LTD
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