Detection reagent for PIEZO2 gene in human gastrointestinal tumor

A detection reagent and gastrointestinal technology, applied in the field of PIEZO2 gene detection reagents in human gastrointestinal tumors, can solve the problems of complex detection process, delay in diagnosis and treatment of patients, long detection time, etc., to achieve flexible and fast detection, targeted Strong and efficient effect

Inactive Publication Date: 2016-10-12
THE AFFILIATED HOSPITAL OF MEDICAL SCHOOL NINGBO UNIV
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Problems solved by technology

However, this invention patent requires PCR detection of 25 pairs of primers on gastrointestinal tumor DNA, which ...

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Embodiment 1

[0010] human gastrointestinal tumors PIEZO2 Gene detection reagent, the detection reagent is a pair of specific primers, the nucleotide sequence of the upstream primer is: 5'- GGAGTTAGGC GGGAGTATAG TAC -3', the nucleotide sequence of the downstream primer is: 5'- TTCTTCAAAA ATAACTAATA CCGAA -3'. is directed at PIEZO2 The fluorescent quantitative methylation-specific amplification upstream primers and fluorescent quantitative methylation-specific amplification downstream primers designed for the promoter region of the gene were confirmed by our extensive screening tests. The primers were provided by Shanghai Shengong Bioengineering Co., Ltd. synthesis.

Embodiment 2

[0012] Utilize a pair of primers of embodiment 1 to carry out case detection, specific detection process is as follows:

[0013] 1. DNA extraction: We collected 5 cases of gastric cancer paraffin samples and corresponding paracancerous paraffin samples from Ningbo Third People's Hospital, and extracted DNA samples with QIAamp DNA FFPE Tissue Kit (purchased from Qiagen, Hilden, Germany). We collected 5 cases of colorectal cancer paraffin samples and corresponding paracancerous tissue paraffin samples from Ningbo Third People's Hospital, and extracted them in the same way to obtain DNA samples. DNA samples require that the ratio of DNA absorbance (A) (A260 / A280) is about 1.80 .

[0014] 2. Methylation of DNA samples: EZ DNA methylation kit (purchased from Zymo, USA) was used to convert the sample DNA to bisulfite, and after the DNA sample was treated with bisulfite, methylation could occur. The cytosine (C) base remains unchanged, and the unmethylated C is converted into uracil...

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Abstract

The invention discloses a detection reagent for a PIEZO2 gene in a human gastrointestinal tumor. The detection reagent comprises a pair of specific primers, wherein the nucleotide sequence of the upstream primer is GGAGTTAGGC GGGAGTATAG TAC, and the nucleotides of the downstream primer is TTCTTCAAAA ATAACTAATA CCGAA. The detection reagent is specific to the PIEZO2 gene in the human gastrointestinal tumor, the relation of the PIEZO2 gene and the tumor is not clear currently, but the fact that in the gastrointestinal tumor, the PIEZO2 gene methylation degree of the PIEZO2 gene of cancer tissue and the PIEZO2 gene methylation degree of the PIEZO2 gene of para-carcinoma tissue, in fluorescent quantitative methylation detection, are different, and the PIEZO2 gene methylation degree of the cancer tissue is apparently higher than that of the para-carcinoma tissue, is discovered. The detection reagent has the advantages of being flexible and quick in detection, simple and convenient, highly targeted, accurate and reliable, high in efficiency and economical, and is beneficial to early detection and immediate treatment of the colorectal cancer and the stomach cancer.

Description

technical field [0001] The invention relates to a detection reagent for digestive system tumors, in particular to human gastrointestinal tumors PIEZO2 Gene detection reagents. Background technique [0002] Digestive system tumors are mainly malignant gastric cancer and colorectal cancer. About half of all deaths from digestive system malignant tumors are caused by gastric cancer. The incidence of colorectal cancer has increased significantly in recent years. The methods currently used to diagnose malignant tumors in the digestive system mainly include digestive endoscopy, imaging examinations, biochemical and immunological examinations, and genetic diagnosis. In the immunological examination, the markers of gastric cancer mainly include CA199, CEA, CA242, CA125, AFP, NES, etc., and the markers of colorectal cancer mainly include CA19-9, CEA, P53, K-ras, etc. Gene diagnosis is a detection technology that has emerged in recent years. It has the advantages of rapidity, specif...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6886C12Q2600/154
Inventor 叶孟李锦芸周重昌倪超杨平陈思毕旭儿
Owner THE AFFILIATED HOSPITAL OF MEDICAL SCHOOL NINGBO UNIV
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