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Gene marker, kit and detection method for detecting pancreas cancer

A gene marker and pancreatic cancer technology, applied in the field of clinical molecular diagnosis of pancreatic cancer, can solve the problems of low sensitivity and specificity of pancreatic cancer, difficulty in widespread and routine application, and difficulty in diagnosing pancreatic tumors, so as to achieve easy dynamic monitoring, High sensitivity and specificity, easy recurrence and metastasis effect

Inactive Publication Date: 2017-11-24
SHANGHAI EPICAN BIOTECH CO LTD +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, imaging is easily affected by operator experience, depends on equipment, and is expensive, especially in the case of limited medical resources. Its accuracy is difficult to guarantee, and it is difficult to be widely and routinely used. CT and ultrasound are difficult to diagnose Pancreatic tumor
Tissue biopsy is currently the gold standard for clinically diagnosing pancreatic cancer, but there are great limitations in tissue biopsy, such as the difficulty of surgical sampling, or the inconvenient puncture of some cancer sites, and the puncture itself will also bring certain clinical risks. Puncture screening will bring great pain to patients
The most widely used serological test is the detection of carcinoembryonic antigen (CEA), but the sensitivity and specificity of CEA for early pancreatic cancer are not high, and it often increases after tumor metastasis

Method used

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  • Gene marker, kit and detection method for detecting pancreas cancer
  • Gene marker, kit and detection method for detecting pancreas cancer
  • Gene marker, kit and detection method for detecting pancreas cancer

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] Example 1. Screening of pancreatic cancer gene markers

[0027] 1) Extract plasma DNA:

[0028] 10 ng of plasma DNA was extracted from samples from 20 pancreatic cancer patients and 20 normal individuals. This step can be carried out using any methods and reagents suitable for extracting plasma DNA well known to those skilled in the art.

[0029] 2) End-fill the plasma DNA, overhang A and connect with the sequencing adapter:

[0030] Prepare a reaction mixture containing 50uL plasma DNA, 7uL End Repair & A-Tailing Buffer and 3uL End Repair & A-Tailing Enzyme mix according to the Kapa Hyper Perp Kit instructions (the total volume is 60uL), incubate at 20°C for 30 minutes, and then incubate Incubate at 65°C for 30 minutes. Prepare the following ligation reaction mixture in a 1.5mL low adsorption EP tube: 5uL Nuclease free water, 30uL Ligation Buffer and 10 uL DNA Ligase. Add 5uL of sequencing adapters to 45uL ligation reaction mixture, mix, heat at 20°C for 20 minutes...

Embodiment 2

[0049] Example 2. Effectiveness of Pancreatic Cancer Gene Markers

[0050] This example verifies the effectiveness of the pancreatic cancer gene markers of the present invention in detecting pancreatic cancer.

[0051] The 5-hmC content of the 10 pancreatic cancer gene markers described in the present invention in the first batch of 82 samples (41 cases of pancreatic cancer and 41 cases of healthy controls) was determined according to the method of Example 1.

[0052] The standardized 5-hmC content of each gene marker is multiplied by the corresponding weighting coefficient of the marker in Example 1 to obtain the predictor t of the gene marker, and then the predictor t of each gene marker is added to obtain The total predictor T, and then the total predictor T is transformed according to the following formula to obtain the score P:

[0053]

[0054] If P>0.5, the subject sample has pancreatic cancer; if P≤0.5, the subject sample is normal.

[0055] figure 1 The results ...

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Abstract

The invention relates to a gene marker, a kit and a detection method for detecting pancreas cancer. The gene marker comprises one or two or more of the following genes: SI, CLEC4C, SNX7, MEOX2, FAT1, FMO3, CFTR, PLPPR3, AFM and COL5A2. The content of 5-hydroxymethylcytosine in the pancreas cancer gene marker is detected through high-throughput sequencing, so as to determine whether the pancreas caner exists. The gene marker for detecting pancreas cancer has the advantages of being safe and noninvasive, having a wide DNA source, being highly accurate, being convenient to operate, and providing good user experience. The gene marker can be combined with other clinical indexes, and provides more accurate determining results for screening, diagnosis, treatment and prognosis of pancreas cancer.

Description

technical field [0001] The present invention relates to the field of clinical molecular diagnosis of pancreatic cancer. Specifically, the present invention relates to a gene marker, a kit and a pancreatic cancer detection method for detecting pancreatic cancer. Background technique [0002] Pancreatic adenocarcinoma (PCA) is a highly malignant gastrointestinal malignancy that is difficult to diagnose and treat. About 90% of them are ductal adenocarcinomas originating from glandular epithelium. The incidence of this disease is higher in men than in women, with a male-to-female ratio of 1.5 to 2:1. Male patients are far more common than premenopausal women, and the incidence of postmenopausal women is similar to that of men. The cause of pancreatic cancer is still unclear. Its occurrence is related to smoking, alcohol consumption, high-fat and high-protein diet, excessive coffee consumption, environmental pollution and genetic factors; recent survey reports have found that t...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
CPCC12Q1/6886C12Q2600/118C12Q2600/154
Inventor 陆星宇宋艳群彭莱张子谋
Owner SHANGHAI EPICAN BIOTECH CO LTD
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