Biomarker for gastric adenocarcinoma diagnosis
A gastric adenocarcinoma and product technology, applied in the field of biomarkers for gastric adenocarcinoma diagnosis, can solve the problems of unsatisfactory sensitivity and specificity, low gastric cancer diagnosis rate, pain and difficulty in risk and so on.
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Embodiment 1
[0078] Example 1 Screening for gene markers associated with gastric adenocarcinoma
[0079] 1. Sample collection
[0080] The paracancerous tissue and cancer tissue samples of 3 patients with gastric adenocarcinoma who underwent radical resection were collected. The patients did not receive any chemotherapy or radiotherapy before gastrectomy. The information of the included patients included gender, age, lauren type, tissue All the specimens mentioned above were obtained with the consent of the organizational ethics committee, and the patients signed an informed consent form.
[0081] 2. Preparation of RNA samples
[0082] Tissue RNA was extracted using QIAGEN tissue RNA extraction kit, and the operation was performed according to the specific steps in the manual.
[0083] Nanodrop2000 was used to detect the concentration and purity of the extracted RNA, agarose gel electrophoresis was used to detect RNA integrity, and Agilent2100 was used to determine the RIN value. Concen...
Embodiment 2
[0103] Example 2 QPCR sequencing to verify the differential expression of related genes
[0104] 1. Large-scale QPCR verification of the differential expression of the LINC00511 gene. According to the sample collection method in Example 1, 10 cases of gastric adenocarcinoma tissue and 10 paracancerous tissue samples were collected.
[0105] 2. The RNA extraction steps are the same as in Example 1.
[0106] 3. Reverse transcription:
[0107] (1) Configuration of reverse transcription system
[0108] Use 25μl reaction system, take 1μg total RNA for each sample as template RNA, and add the following components to PCR tubes: DEPC water, 5× reverse transcription buffer, 10mM dNTP, 0.1mM DTT, 30μM Oligo dT, 200U / μl M-MLV, template RNA. Incubate at 42°C for 1 hour, then centrifuge briefly at 72°C for 10 minutes.
[0109] (2) QPCR amplification test
[0110] Primer design
[0111] The primer sequence of LINC01234 gene is:
[0112] Forward primer: 5'-CGTGAAGAGTAGATGTAGA-3' (SEQ...
Embodiment 3
[0124] Example 3 Analysis of the expression of LINC01234 in the TCGA database
[0125] 1. Data collection
[0126] The lncRNA expression profile data of 285 cases of gastric adenocarcinoma tissues and 30 cases of adjacent tissues were collected from the TCGA database, and the expression level of lncRNA LINC01234 in gastric adenocarcinoma tissues and adjacent tissues was analyzed; box plots were drawn.
[0127] 2. ROC curve analysis
[0128] The receiver operating characteristics of lncRNALINC01234 were analyzed using the pROC package in R language, the binomial exact confidence space was calculated, and the ROC curve was drawn.
[0129] 3. Results
[0130] The expression level of LINC01234 as figure 2 As shown, compared with the control group, the expression of LLINC01234 (P=6.78e-16) was significantly up-regulated in gastric adenocarcinoma tissue.
[0131] The ROC curve of LINC01234 is as image 3 As shown, the AUC value of LINC01234 is as high as 0.894, and has high sp...
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