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Primer for detecting peripheral blood exosome circ-LRP8 and kit, and application thereof in gastric cancer liver metastasis

A technology of peripheral blood and exosomes, applied in the determination/testing of microorganisms, biochemical equipment and methods, DNA/RNA fragments, etc.

Active Publication Date: 2020-08-21
江苏华美健晟生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

It has been reported that CD44v6, OPN, YB-1, MAGE-A10 and other molecules may be used as potential markers of gastric cancer liver metastasis, but further verification is still needed

Method used

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  • Primer for detecting peripheral blood exosome circ-LRP8 and kit, and application thereof in gastric cancer liver metastasis
  • Primer for detecting peripheral blood exosome circ-LRP8 and kit, and application thereof in gastric cancer liver metastasis
  • Primer for detecting peripheral blood exosome circ-LRP8 and kit, and application thereof in gastric cancer liver metastasis

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] Example 1 Detection of circ-LRP8 in peripheral blood exosomes

[0030] 1. Obtain peripheral blood samples to be tested

[0031] Peripheral blood samples from 120 cases of gastric cancer and 120 cases of normal subjects.

[0032] The samples are the peripheral blood samples of the patients with gastric cancer and normal people with coagulant added.

[0033] 2. Extract exosomes from peripheral blood samples to be tested and verify

[0034] Extract exosomes from gastric peripheral blood 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)...

Embodiment 2

[0063] Example 2 Clinical significance of peripheral blood exosome circ-LRP8

[0064] The correlation between the expression of peripheral blood exosomal RNA hsa_circ-LRP8 and clinical parameters such as gender, age, tumor diameter, TNM stage, degree of tissue differentiation, lymph node metastasis, and liver metastasis of gastric cancer was further analyzed.

[0065] Table 4 Clinical significance analysis of peripheral blood exosome circ-LRP8

[0066]

[0067] Table 5 Results of multivariate analysis

[0068]

[0069]

[0070] It can be seen from Table 4 and Table 5 that the expression level of peripheral blood exosomal RNA hsa_circ-LRP8 was significantly correlated with the depth of invasion, lymph node metastasis, liver metastasis, and TNM stage (P values ​​were 0.002, 0.001, 0.001, and 0.001, respectively), while the There was no significant correlation between gender, age, tumor location, tumor diameter and tissue differentiation (P values ​​were 0.852, 0.709, 0...

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Abstract

The invention relates to a primer for detecting peripheral blood exosome circ-LRP8, a kit, a detection method and an application thereof. The kit comprises a peripheral blood exosome RNA extraction reagent, the primer, a genome-removed DNA reaction system, a reverse transcription reaction system and a qPCR operation reaction system. The invention discloses an expression condition of circ-LRP8 in peripheral blood exosomes of gastric cancer patients and normal people, and the circ-LRP8 in the peripheral blood exosomes is detected by using fluorescent quantitative PCR. The circ-LRP8 can be used for early diagnosis of liver metastasis of gastric cancer, judging disease progression and prognosis conditions and evaluating the treatment effect of liver metastasis of gastric cancer.

Description

technical field [0001] The invention relates to a primer and a kit for detecting peripheral blood exosome circ-LRP8 and its application in liver metastasis of gastric cancer, belonging to the technical field of molecular diagnosis. Background technique [0002] Gastric cancer is a serious threat to human health and is one of the five major cancers in the world. The World Health Organization statistics report shows that in 2012, there were about 951,000 new cases of gastric cancer worldwide, and about 723,000 deaths. Gastric cancer rates are higher in East Asia (particularly China, Korea, Mongolia, and Japan), Central and Eastern Europe, South America, and most of Africa. China is the country with the highest incidence and mortality rate of gastric cancer in the world. According to data from the National Cancer Registry Center, there were about 679,000 new cases of gastric cancer in China in 2015, and about 498,000 deaths, ranking second among malignant tumors in my country....

Claims

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Application Information

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IPC IPC(8): C12Q1/6886C12N15/113C12N15/11
CPCC12Q1/6886C12Q2600/158C12Q2600/178C12Q2600/118C12Q2600/106
Inventor 邹晨范钰郎亚昆吴朝晖朱宝平
Owner 江苏华美健晟生物科技有限公司
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