Micro RNA (miRNA) biomarker combination and kit used for diagnosing or assisting in diagnosing gastric cancer
A technology for biomarkers and auxiliary diagnosis, which is applied in the direction of biochemical equipment and methods, microbiological determination/inspection, etc., can solve problems such as missed diagnosis, and achieve the effect of excellent accuracy and sensitivity
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Embodiment 1
[0059] Eleven significantly up-regulated miRNAs in gastric cancer tissues were screened out through the TCGA high-throughput miRNA expression database (n=477), including miR-21, miR-196a-1, miR-146b, miR-196b, miR-135b, miR -181b-1, miR-181a-1miR-93, miR-335, miR-196a-2 and miR-18a, 10 miRNAs after cross-platform verification, miR-21, miR-196a, miR-146b, miR -196b, miR-135b, miR-181b-1, miR-181a-1miR-93, miR-335 and miR-18a, wherein miR-196a-1 and miR-196a-2 are the same sequence (miR-196a) , please refer to Table 1 for the specific sequence.
[0060] Table 1 Sequences of miRNA biomarker combinations
[0061]
[0062] The prediction effects of the above 10 miRNAs were verified on two independent high-throughput data sets (the screening characteristics of the data sets refer to Table 5). For the results, please refer to the attached figure 1 , the area under the ROC curve AUC obtained was greater than 0.93.
[0063] Based on the relationship between the expression of miRN...
Embodiment 2
[0068] The miRNA biomarker combination provided in Example 1 was used for screening of early gastric cancer.
[0069] 1. Serum Sample Collection
[0070] Whole blood was collected in Sarstedt with coagulant Serum-Gel 9ml tubes (cat.no.02.1388) (or other collection tubes without coagulant or EDTA). For complete coagulation, the test tube containing the coagulation accelerator needs to be placed at room temperature (15-25°C) for 10 minutes to 1 hour. It should be noted that the test tube containing the coagulant needs to be placed for 10 minutes, and the test tube without the coagulant needs to be placed for at least 30 minutes;
[0071] Centrifuge the test tube at 1900×g (3000rpm), 4°C for 10 min; transfer the supernatant serum (yellow) to a new test tube with a conical bottom (usually 10 ml of whole blood can obtain 3-5 ml of serum); put the conical bottom The plasma in the test tube was centrifuged at 16,000×g at 4°C for 10 minutes; the supernatant was transferred to a ne...
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