Gene marker for detecting esophageal cancer and use thereof and detection method

A gene marker and esophageal cancer technology, applied in the field of clinical molecular diagnosis of esophageal cancer, can solve the problems of lack of simple and effective

Pending Publication Date: 2018-06-01
SHANGHAI EPICAN BIOTECH CO LTD +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the existing technology lacks simple and effective diagnostic methods, especially suitable for population census, so it is of great significance to find new early diagnostic markers for esophageal cancer

Method used

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  • Gene marker for detecting esophageal cancer and use thereof and detection method
  • Gene marker for detecting esophageal cancer and use thereof and detection method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] Example 1: Screening of esophageal cancer gene markers:

[0034] (1) Extraction of plasma DNA: 10 ng of plasma DNA was extracted from samples from 20 patients with esophageal cancer and 20 normal subjects. This step can be carried out using any methods and reagents suitable for extracting plasma DNA well known to those skilled in the art.

[0035] (2) End-fill the plasma DNA, overhang A and connect with the sequencing adapter:

[0036] 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 30min, then incubate at 65°C ℃ warm bath for 30min. 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. 5uL of sequencing adapters were added to 45uL of the ligation reaction mixture, mixed, heated at 20°C for 20...

Embodiment 2

[0052] Example 2: Effectiveness of esophageal cancer gene markers:

[0053] This example is used to verify the effectiveness of the esophageal cancer gene markers of the present invention in detecting esophageal cancer. According to the method of Example 1, the 5-hmC content of the 10 esophageal cancer gene markers of the present invention in the first batch of 82 samples (41 cases of esophageal cancer and 107 cases of healthy controls) was determined. 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 obtained by Logistic transformation according to the following formula to obtain the score P: P=1 / (1+e -T ), if P>0.5, the subject sample has esophageal cancer; if P≤0.5, the subject sample is normal. figure 1 The results of disti...

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Abstract

The invention discloses a gene marker for detecting esophageal cancer and a use thereof and a detection method. The gene marker comprises one or more of genes of a phospholipid inositol phosphatase CXdomain containing protein 3, a sulfatase 1, a F-Box and leucine-rich repeat protein 7, a Tolloid-like protein 1, a RNA binding template single-stranded function protein 3, an ADAMTS-like protein 1, phosphodiesterase 10A, a LIM-domain ligase 2, a gamma-aminobutyric acid type A receptor protein in the gamma 3 subfamily and a sequence similar 155 family protein A. The gene marker is used for detecting esophageal cancer. The gene marker is suitable for a more extensive source of DNA samples, the detection is safe and non-invasive, the asymptomatic people have high acceptance to detection, the accuracy is high, the gene marker has high sensitivity and specificity to the early esophageal cancer, the operation is convenient, the user experience is good and the dynamic monitoring of the recurrence and metastasis of esophageal cancer is easy.

Description

technical field [0001] The invention belongs to the technical field of clinical molecular diagnosis of esophageal cancer, in particular, the invention relates to a method and a reagent for detecting the presence of esophageal cancer by detecting the 5-hydroxymethylcytosine content of esophageal cancer gene markers through high-throughput sequencing box. Background technique [0002] Esophageal cancer, as one of the cancers with the highest incidence of malignant tumors in my country, has been paid more and more attention by people. There are many pathogenesis factors for esophageal cancer. It is currently recognized that drinking and smoking are various chronic stimuli that cause damage to the esophagus, and environmental factors are the main causes of esophageal squamous cell carcinoma in China. The survey found that eating hot food, excessive alcohol consumption, low income, low body mass index, previous esophageal lesions, not eating on time, eating spicy food and family...

Claims

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Application Information

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