Application of cystatin SN and AFP in preparation of marker for diagnosis and predicting of 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 easy portability, good repeatability, and convenient use

Active Publication Date: 2014-07-23
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 SN and AFP in preparation of marker for diagnosis and predicting of liver cancer
  • Application of cystatin SN and AFP in preparation of marker for diagnosis and predicting of liver cancer
  • Application of cystatin SN and AFP in preparation of marker for diagnosis and predicting of liver cancer

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

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

[0025] Coat the ELISA plate with a mouse anti-human Cystatin SN monoclonal antibody at a concentration of 5 μg / mL, coat it overnight at 4°C, and wash the plate; then block it in 2% BSA at room temperature for 2 hours, and wash the plate; Then concentration is 0pg / mL, 50pg / mL, 100pg / mL, 200pg / mL, 400pg / mL, 800pg / mL, the Cystatin SN protein standard substance of 1600pg / mL (the aminoacid sequence of coding Cystatin SN protein is as SEQ ID NO. 1) and serum samples were added to the blocked plate, reacted at 37°C for 1 hour, and washed the plate; then reacted with rabbit anti-human Cystatin SN polyclonal antibody with a concentration of 0.5 μg / mL HRP at 37°C for 1 hour, 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 ). Depend on figure 1 It can be seen that the...

Embodiment 2

[0032]Embodiment 2 Cystatin SN enzyme-linked immunoassay kit

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

[0034] Table 1. Cystatin SN ELISA kit components

[0035]

[0036]

[0037] Evaluation of Cystatin SN ELISA kit: use Cystatin SN ELISA kit to detect Cystatin SN positive quality control products, and repeat the detection 10 times at two levels of Cystatin SN protein concentration of 160pg / mL and 80pg / mL respectively, the 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 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 3A

[0038] Example 3 AFP enzyme-linked immunosorbent assay (ELISA) detection kit

[0039] The AFP-ELISA detection kit uses the product of Beijing Rejing Biotechnology Co., Ltd. (Cat. No. 1003). The amino acid sequence of the detected AFP marker is shown in SEQ ID NO.2.

[0040] The Cystatin SN enzyme-linked immunoassay kit constructed in Example 2 was combined with the AFP-ELISA detection kit in this example to form a Cystatin SN-AFP combined detection kit.

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Abstract

The invention discloses a combined application of a cystatin SN and an alpha-fetoprotein (AFP), and particularly discloses the application of the cystatin SN and the AFP in preparation of a marker for diagnosis and predicting of liver cancer. The invention also discloses a capture agent of the liver cancer marker. A liver cancer detection kit is formed by combining the capture agent with conventional reagents. The obtained kit has the advantages of good specificity, high sensitivity and the like, can be used for early diagnosis of liver cancer, therapeutic effect evaluation in a therapeutic process and monitoring of metastasis and recurrence after therapy, and has test results sooner than clinical symptoms.

Description

technical field [0001] The invention belongs to the field of medical detection and relates to the application of Cystatin SN and AFP in the preparation of 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. HCC incidence and mortality rates are almost identical, predicting poor overall survival for HCC. 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 factor for the development of liv...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/574G01N33/577G01N33/68
CPCG01N33/57438G01N33/57484G01N2333/471G01N2333/8139G01N2800/52G01N2800/54
Inventor 王弢秦勇吴孝林
Owner SHANGHAI LIANGRUN BIOMEDICINE TECH CO LTD
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