Primer and reagent kit for detecting serum exosome RP11-439M11-1, and application of primer or reagent kit to diagnosis and treatment of gastric cancer liver metastasis
A technology of exosomes and kits, which is used in the determination/inspection of microorganisms, biochemical equipment and methods, DNA/RNA fragments, etc.
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Embodiment 1
[0035] Example 1 Detection of RP11-439M11-1 in serum exosomes
[0036] 1. Obtain the serum sample to be tested:
[0037] Serum samples from 120 cases of gastric cancer and 120 normal persons.
[0038] The samples are the serum samples of the patients with gastric cancer and normal people with coagulant added.
[0039] 2. Extract exosomes from serum samples to be tested and verify
[0040] Extract exosomes from gastric serum according to the instructions of the exosome extraction kit, and the steps are as follows: (1) Whole blood is centrifuged at 3000×g for 15 min (centrifugal radius 10 cm) to remove cells or cell debris. (2) Put the upper liquid into a centrifuge tube, add 63 μl ExoQuick reagent to 250 μl serum, put it at 4°C for 30 minutes; (3) Centrifuge the mixture at 1500×g (centrifugal radius 10cm) for 30 minutes at 4°C; ( 4) Aspirate all the supernatant, centrifuge at 1500×g (centrifugal radius 10cm) for 5 min at 4°C, and aspirate the entire supernatant; (5) Dissolve...
Embodiment 2
[0071] Example 2 Clinical significance of serum exosome RP11-439M11-1
[0072] The correlation between serum exosomal RP11-439M11-1 expression and clinical parameters such as gender, age, tumor diameter, clinical stage, degree of tissue differentiation, lymph node metastasis, and liver metastasis of gastric cancer was further analyzed. See Table 4 and Table 5. It was found that the expression level of serum exosomal RP11-439M11-1 was significantly correlated with the depth of invasion, lymph node metastasis, liver metastasis, and clinical stage (P values were 0.006, 0.028, 0.001, and 0.001, respectively). There was no significant correlation with gender, age, tumor location, tumor diameter, tissue differentiation, Helicobacter pylori infection and CEA level. The results of multivariate analysis suggested that the high expression of serum exosomal RP11-439M11-1 was one of the independent prognostic factors for gastric cancer liver metastasis.
[0073] Table 4 Clinical signif...
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