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A marker gene cst1 and its application

A gene and gallbladder polyp technology, applied in biochemical equipment and methods, material inspection products, instruments, etc., can solve the problems of misdiagnosis and mistreatment of multiple polyps, and achieve the effect of convenient clinical use and ensuring consistency

Active Publication Date: 2015-12-30
QINGDAO MEDINTELL BIOMEDICAL CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Misdiagnosis of multiple polyps in clinic due to multiple stones, cholecystectomy for fear of polyp malignant transformation, resulting in mistreatment

Method used

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  • A marker gene cst1 and its application
  • A marker gene cst1 and its application
  • A marker gene cst1 and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] Example 1 Collection of research samples and RNA extraction

[0028] Collect 7 samples of gallbladder stones and 2 samples of gallbladder polyps for RNA extraction. After RNA extraction, agarose gel electrophoresis can be used to preliminarily determine whether the quality of the extracted RNA samples is qualified or not, and whether they can be used for further transcriptome analysis . Then, the extraction of RNA samples was detected by NanoDrop1000 spectrophotometer, and the sample requirements for RNA-seq sequencing: OD260 / OD280 was 1.8-2.2.

Embodiment 2

[0029] Embodiment 2 data analysis

[0030]Sequencing platform: Illumina's HiSeq2500 high-throughput sequencing platform

[0031] After high-throughput transcriptome deep sequencing, in order to better understand the functions of differentially expressed genes, we enriched the differentially expressed genes with GeneOnlogy and KEGG pathways, and performed functional annotation and protein interaction network analysis on the differentially expressed genes. In view of the results of the above data analysis, combined with the literature, we screened a differentially expressed gene CST1 that was down-regulated in gallbladder polyps and up-regulated in gallbladder stones.

Embodiment 3

[0032] Example 3 CST1 gene expression in gallbladder polyps and gallbladder stones

[0033] 1. Materials and methods

[0034] 1. Materials

[0035] Gallbladder polyps and gallbladder stones were collected from 45 and 38 inpatients from 2005 to 2010, respectively.

[0036] 2. Method

[0037] 2.1 Extraction of total RNA from gallbladder polyps and gallbladder stones

[0038] Extract RNA from gallbladder tissue and paratumor tissue according to the instructions of Kangwei Century Ultrapure RNA Extraction Kit (UltrapureRNAKit (DNaseI), Cat. No. CW0597), prove the integrity of RNA by gel electrophoresis, and measure the concentration and purity of RNA with a nucleic acid protein analyzer .

[0039] Use the Ultrapure RNA Extraction Kit (Cat. No. CW0597) to extract, and the main steps are as follows:

[0040] 1. Sample processing, 30-50mg tissue is fully ground in liquid nitrogen and then added 1ml BufferRLT, or homogenized after adding 1ml BufferRLT to the tissue sample.

[00...

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Abstract

The invention relates to a marker gene CST1 and application thereof. High throughput transcriptome depth sequencing and bioinformatics analysis are carried out on gallbladder polyp and gallstone tissues, one gene which is down-regulated in gallbladder polyp and up-regulated in gallstone is screened out, further classical molecular biological experimental verification is carried out by adopting fluorogenic quantitative PCR (polymerase chain reaction), and results show that the screened CST1 gene can be taken as a specific labelled molecule used for distinguishing the gallbladder polyp from the gallstone. The marker gene provided by the invention can be applied to preparation of an auxiliary diagnosis preparation used for distinguishing the gallbladder polyp from the gallstone and has important clinical application value.

Description

technical field [0001] The invention relates to a marker gene and its application. Specifically, the present invention relates to a marker gene CST1 (cystatinSN) and its application in the preparation of auxiliary diagnostic preparations for distinguishing gallbladder polyps and gallbladder stones. Background technique [0002] Gallbladder stone (cholelithiasis) is one of common disease in my country, and the prevalence rate of natural population is about 10% above, can cause serious consequence, as severe biliary tract infection, obstructive jaundice, biliary pancreatitis, and biliary tract tumor also are closely related and threaten people's health. The causes of gallstone formation are complex, and can be divided into three types of gallstones according to their components: cholesterol stones, pigment stones, and mixed stones. For a long time, many scholars have devoted themselves to the study of the pathogenesis of cholesterol gallstones. Stone-causing genes, genetic e...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/68G01N33/68
CPCC12Q1/6883C12Q2600/158G01N33/6893G01N2800/08
Inventor 杨承刚常鹏孙锦云李红伟
Owner QINGDAO MEDINTELL BIOMEDICAL CO LTD