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Application of Cystatin S and CA15-3 in preparation of markers for diagnosing and predicting breast cancer

A marker and breast cancer technology, which is applied in the field of medical detection, can solve the problems of increasing the complexity of treatment and not being able to diagnose breast cancer, and achieve the effects of improving sensitivity and specificity, being easy to carry, and having good repeatability

Active Publication Date: 2014-07-02
SHANGHAI LIANGRUN BIOMEDICINE TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Methods such as biopsy and imaging examinations have high diagnostic accuracy, but when patients are diagnosed, they are often in the middle and advanced stages, which increases the complexity of treatment
At present, many studies have found that serum tumor markers can be used in the diagnosis of tumors. The existing technology discloses that carbohydrate antigen 15-3 (CA15-3) is used as a tumor marker to diagnose breast cancer, but there are false positives in the diagnosis of a single marker, and it cannot for the diagnosis of early breast cancer

Method used

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  • Application of Cystatin S and CA15-3 in preparation of markers for diagnosing and predicting breast cancer
  • Application of Cystatin S and CA15-3 in preparation of markers for diagnosing and predicting breast cancer
  • Application of Cystatin S and CA15-3 in preparation of markers for diagnosing and predicting breast cancer

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] Embodiment 1 establishes Cystatin S serum detection system and its optimization

[0024] Coat the ELISA plate with a mouse anti-human Cystatin S monoclonal antibody at a concentration of 5 μg / mL, and the coating condition is to coat overnight at 4°C, wash the plate; then block in 2% BSA at room temperature for 2 hours , wash the plate; the Cystatin S protein standard substance (the amino acid sequence of encoding Cystatin S such as SEQ ID NO.1) and samples were added to the closed plate, reacted at 37°C for 1 hour, and washed the plate; then reacted with rabbit anti-human Cystatin S polyclonal antibody labeled with HRP at a concentration of 0.5 μg / mL at 37°C for 1 hour , wash the plate; then react with tetramethylbenzidine (TMB) for 2-3 minutes, and finally stop the reaction with 2M sulfuric acid, and detect the OD value under the condition of 450nm ( figure 1 ). The results showed that the linear range of Cystatin S was 50pg / mL-1600pg / mL, the linear correlation coeff...

Embodiment 2

[0031] Embodiment 2 Cystatin S ELISA kit

[0032] According to the establishment of the Cystatin S serum detection reaction system in Example 3, the Cystatin S enzyme-linked immunoassay kit was constructed, and the specific components are as shown in Table 1:

[0033] Table 1. Cystatin S ELISA kit components

[0034]

[0035]

[0036] 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 two levels of Cystatin S protein concentration of 160pg / mL and 80pg / mL, and the test results show Coefficient of variation CV ≤ 10%; use 3 batches of kits to detect the same sample, and the coefficient of variation between batches of 3 batches of kits CV ≤ 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 3CA153

[0037] Embodiment 3CA153 ELISA kit

[0038] CA153 enzyme-linked immunoassay kit was purchased from Beijing Rejing Biotechnology Co., Ltd. The amino acid sequence of the detected CA153 marker is shown in SEQ ID NO.2.

[0039] Combine the kit purchased in this example with the Cystatin S enzyme-linked immunoassay kit constructed in Example 2 to form a Cystatin S-CA153 combined detection kit, which is used for the diagnosis and prognostic detection of breast cancer.

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Abstract

The invention discloses combined application of cysteine proteinase inhibitor S (Cystatin S) and carbohydrate antigen 15-3 (CA15-3) and in particular discloses application of Cystatin S and CA15-3 in preparation of markers for diagnosing and predicting breast cancer. The invention also discloses a trapping agent of breast cancer markers and a kit comprising the trapping agent. The kit has the advantages of good specificity and high sensitivity and can be applied to early diagnosis of the breast cancer, therapeutic effect evaluation in a breast cancer treating process and monitoring of transfer and recurrence after treating the breast cancer; diagnosis results can be earlier than clinical symptoms.

Description

technical field [0001] The invention belongs to the field of medical detection, and relates to the application of cysteine ​​protease inhibitor S (Cystatin S) and carbohydrate antigen 15-3 (CA15-3) in preparing markers for diagnosing and predicting breast cancer. Background technique [0002] Breast cancer is one of the most common malignant tumors in women, and its incidence rate is increasing year by year. It ranks first in the incidence and death of women in the world, and has become a major threat to women's health. According to the International Agency for Research on Cancer (IARC) report, in 2008, there were 1.38 million new cases of breast cancer in women worldwide, accounting for 22.9% of female malignant tumors, and 460,000 deaths, accounting for 13.7% of female malignant tumor deaths. %. The 2010 "China Breast Disease Survey Report" pointed out that the growth rate of the incidence of breast cancer in China is 1 to 2 percentage points higher than that of Western c...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68G01N33/574
CPCG01N33/57415G01N2800/365G01N2800/52G01N2800/7028
Inventor 王弢秦勇渠香云
Owner SHANGHAI LIANGRUN BIOMEDICINE TECH CO LTD
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