Early diagnosis reagent for colorectal cancer based on combined detection of methylation levels of SDC2 gene and SFRP2 gene
A colorectal cancer, 1.SDC2 technology, applied in the field of molecular biology, can solve the problems of low accuracy, time-consuming and laborious, invasive and so on
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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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