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Primers and kit for detecting serum exosome circ-KCNQ1 and application of primers and kit in gastric cancer with liver metastasis

A technology for detection kits and detection primers, which can be used in recombinant DNA technology, microbial measurement/testing, biochemical equipment and methods, etc., and can solve the problem of low specificity

Active 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

Some tumor molecular markers have been used in clinical early detection and auxiliary diagnosis, such as carcinoembryonic antigen CEA, CA199, CA125, AFP, etc., but the specificity is still low.

Method used

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  • Primers and kit for detecting serum exosome circ-KCNQ1 and application of primers and kit in gastric cancer with liver metastasis
  • Primers and kit for detecting serum exosome circ-KCNQ1 and application of primers and kit in gastric cancer with liver metastasis
  • Primers and kit for detecting serum exosome circ-KCNQ1 and application of primers and kit in gastric cancer with liver metastasis

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] Example 1 Detection of circ-KCNQ1 in serum exosomes

[0031] 1. Obtain the serum sample to be tested:

[0032] Serum samples from 120 cases of gastric cancer and 120 normal persons.

[0033] The samples are the serum samples of the patients with gastric cancer and normal people with coagulant added.

[0034] 2. Extract exosomes from serum samples to be tested and verify

[0035] 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 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 radius 10cm) for 5 min at 4°C, and aspirate the entire supernatant; (5) Disso...

Embodiment 2

[0064] Example 2 Clinical significance of serum exosome circ-KCNQ1

[0065] The correlation between serum exosomal hsa_circ-KCNQ1 expression and clinical parameters such as gender, age, tumor diameter, TNM stage, tissue differentiation, lymph node metastasis, and distant metastasis of gastric cancer was further analyzed.

[0066] Table 4 Clinical significance of serum exosomal circ-KCNQ1

[0067]

[0068] Table 5 Multivariate Analysis Results

[0069]

[0070] It can be seen from Table 4 and Table 5 that the expression level of hsa_circ-KCNQ1 in serum exosomes was significantly correlated with the depth of gastric cancer tissue invasion, lymph node metastasis, liver metastasis, and TNM stage (P values ​​were 0.012, 0.021, 0.004, and 0.002, respectively), while the expression level of its patients Gender, age, tumor location, tumor diameter and tissue differentiation were not significantly correlated (P values ​​were 0.707, 0.855, 0.966, 0.201 and 0.435, respectively). ...

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Abstract

The invention relates to detection primers and a detection kit for circ-KCNQ1 in serum exosome and application of the detection primers and the detection kit. The detection kit comprises an RNA extraction reagent of a serum 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 exosome circ-KCNQ1 in serum exosomes of gastric cancer patients and normal people, and the circ-KCNQ1 is detected by using fluorescent quantitative PCR. The serum exosome circ-KCNQ1 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 serum exosome circ-KCNQ1 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 (WHO) 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 the data of the National Central Cancer Registry of China (NC Gastric Cancer), there were about 679,000 new cases of gastric cancer in China in 2015, and about 498,000 deaths, ranking second among mal...

Claims

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Application Information

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IPC IPC(8): C12Q1/6886C12N15/113C12N15/11
CPCC12Q1/6886C12Q2600/118C12Q2600/158C12Q2600/178
Inventor 吴朝晖范钰朱宝平
Owner 江苏华美健晟生物科技有限公司
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