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Primers and kit for detecting peripheral blood exosome circc-PPCDC and application of primer and kit in gastric cancer with liver metastasis

A peripheral blood, exosome technology, applied in recombinant DNA technology, microbial assay/test, biochemical equipment and methods, etc.

Pending Publication Date: 2020-08-18
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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  • Primers and kit for detecting peripheral blood exosome circc-PPCDC and application of primer and kit in gastric cancer with liver metastasis
  • Primers and kit for detecting peripheral blood exosome circc-PPCDC and application of primer and kit in gastric cancer with liver metastasis
  • Primers and kit for detecting peripheral blood exosome circc-PPCDC and application of primer and kit in gastric cancer with liver metastasis

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] Example 1 Detection of circ-PPCDC in peripheral blood exosomes

[0036] 1. Obtain the peripheral blood sample to be tested:

[0037] Peripheral blood samples from 120 cases of gastric cancer and 120 cases of normal subjects.

[0038] The samples are the peripheral blood samples of the patients with gastric cancer and normal people with coagulant added.

[0039] 2. Extract exosomes from peripheral blood samples to be tested and verify

[0040] 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 layer 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 r...

Embodiment 2

[0068] Example 2 Clinical significance of peripheral blood exosome circ-PPCDC

[0069] The correlation between the expression of circ-PPCDC in peripheral blood 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.

[0070] Table 4 Clinical significance of peripheral blood exosome circ-PPCDC

[0071]

[0072]

[0073] Table 5 Multivariate Analysis Results

[0074]

[0075] From Table 4 and Table 5, it can be seen that the expression level of circ-PPCDC in peripheral blood exosomes was significantly correlated with the depth of invasion, lymph node metastasis, liver metastasis, and TNM stage (P values ​​were 0.018, 0.023, 0.016, and 0.015), while that of patients with Gender, age, tumor location, tumor diameter and tissue differentiation were not significantly correlated (P values ​​were 0.707, 0.584, 0.801, 0.100 and 0.215, respectivel...

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Abstract

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

Description

technical field [0001] The invention relates to a primer and a kit for detecting peripheral blood exosome circ-PPCDC 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): C12Q1/6886C12N15/11
CPCC12Q1/6886C12Q2600/118C12Q2600/158C12Q2600/178
Inventor 吴朝晖范钰郎亚昆邹晨朱宝平
Owner 江苏华美健晟生物科技有限公司
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