Application of Cystatin S and AFP to prepare marker for diagnosing and indicating liver cancer

A technology for markers and liver cancer, applied in the field of medical detection, can solve the problem that the specificity and sensitivity of liver cancer cannot meet the monitoring of liver cancer, and achieve the effect of high sensitivity, good repeatability, and convenient use

Active Publication Date: 2014-07-16
SHANGHAI LIANGRUN BIOMEDICINE TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Alpha-fetoprotein (AFP) is the most commonly used tumor molecular marker for monitoring liver cancer worldwide, but the proportion of AFP-negative liver cancer in new cases is increasing, and the specificity and sensitivity of AFP in diagnosing liver cancer can no longer meet the role of monitoring liver cancer

Method used

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  • Application of Cystatin S and AFP to prepare marker for diagnosing and indicating liver cancer
  • Application of Cystatin S and AFP to prepare marker for diagnosing and indicating liver cancer
  • Application of Cystatin S and AFP to prepare marker for diagnosing and indicating liver cancer

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] Example 1 Establishment of Cystatin S Serum Detection Reaction System and Its Optimization

[0025] Coat the ELISA plate with a mouse anti-human Cystatin S monoclonal antibody at a concentration of 5 μg / mL, coat it overnight at 4°C, and wash the plate; then block it in BSA with a mass fraction of 2% at room temperature for 2 hours, and wash the plate; Cystatin S protein standards with concentrations of 0pg / mL, 50pg / mL, 100pg / mL, 200pg / mL, 400pg / mL, 800pg / mL, and 1600pg / mL (the amino acid sequence encoded by Cystatin S is shown in SEQ ID NO.1) Add the samples into the closed plate, react at 37°C for 1 hour, and wash the plate; then use the rabbit anti-human Cystatin S polyclonal antibody labeled with HRP at a concentration of 0.5 μg / mL, react at 37°C for 1 hour, and wash the plate; React with tetramethylbenzidine (TMB) for 2-3 minutes, and finally stop the reaction with 2M sulfuric acid, and detect the OD value at 450nm ( figure 1 ). The results showed that the linear ...

Embodiment 2

[0032]Example 2 Cystatin S ELISA Kit

[0033] According to the Cystatin S serum detection system established in Example 3, the Cystatin S enzyme-linked immunoassay kit was constructed, and the specific components are as shown in Table 1:

[0034] Table 1. Cystatin S ELISA kit components

[0035]

[0036]

[0037] Evaluate the Cystatin S ELISA kit: use the Cystatin S ELISA kit to detect the Cystatin S positive quality control product, repeat the detection 10 times at the levels of Cystatin S protein concentrations of 160pg / mL and 80pg / mL2, and the test results It shows that the coefficient of variation CV≤10%; if the same sample is tested with 3 batches of kits, the inter-assay coefficient of variation CV of the 3 batches of kits is ≤15%. The research on the stability of the kit shows that it can be kept stable after being stored at 4°C for 8 months, stored at 4°C for 2 months after opening, and transported at 0-4°C for 7 days.

Embodiment 3

[0038] Embodiment 3 establishes AFP ELISA kit

[0039] AFP enzyme-linked immunoassay kit was purchased from Beijing Rejing Biotechnology Co., Ltd. (Product No.: 1003).

[0040] The AFP enzyme-linked immunosorbent assay kit and the Cystatin S enzyme-linked immunosorbent assay kit constructed in Example 2 form a Cystatin S-AFP combined detection kit, which is used to detect liver cancer tumor markers AFP and Cystatin S, and the detected AFP marker The amino acid sequence is shown in SEQ ID NO.2.

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Abstract

The invention discloses combined application of Cystatin S and alpha fetoprotein (AFP), and specifically discloses application of Cystatin S and AFP to prepare a marker for diagnosing and indicating liver cancer. The invention also discloses a trapping agent of the liver-cancer marker, and a liver-cancer detection kit is formed by combining the trapping agent and routine reagents. The kit has the advantages of good specificity, high sensitivity and the like, is applicable to early diagnosis on liver cancer, assessment on treatment effect during treatment and monitoring on metastasis and recurrence after treatment is finished, and the diagnosis result is earlier than clinic symptoms.

Description

technical field [0001] The invention belongs to the field of medical detection and relates to the application of Cystatin S and AFP in preparing markers for diagnosis and prediction of liver cancer. Background technique [0002] Liver cancer is the fifth most common tumor in the world. According to the report of the World Health Organization, the growth rate of liver cancer mortality is the second fastest among all cancers. More than 500,000 people worldwide suffer from liver cancer every year, and more than half of them are in China. The incidence and mortality of liver cancer are almost the same, indicating that the overall survival of liver cancer is relatively poor. Therefore, finding appropriate tumor molecular markers to achieve early diagnosis of liver cancer, efficacy evaluation, and recurrence monitoring is of great value in reducing liver cancer mortality and improving the survival status of liver cancer patients. [0003] Liver cirrhosis is an important risk fac...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68G01N33/577G01N33/574G01N33/543
CPCG01N33/57488G01N33/68
Inventor 王弢秦勇渠香云
Owner SHANGHAI LIANGRUN BIOMEDICINE TECH CO LTD
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