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Combined application of cystatin SN and carcino-embryonic antigen

A cysteine ​​protease, carcinoembryonic antigen technology, applied in the field of medical detection, can solve the problems of poor accuracy, poor prognosis, no detection of gastric cancer or gastrointestinal stromal tumor, etc., and achieves convenient use, portability and repeatability. Good results

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

The clinical significance of CEA detection in the serum of patients with gastric cancer is: (1) It is related to advanced poorly differentiated adenocarcinoma—it is also related to tumor size, serosa surface invasion and lymphatic metastasis; (2) It can mediate the contact between tumor cells and liver parenchyma (3) It can be used in combination with other indicators, such as combined with cytology in peritoneal lavage fluid to indicate peritoneal recurrence and peritoneal implantation of gastric cancer, but the accuracy is worse than CA125; (4) It can be combined with other indicators It can be used to evaluate the efficacy of chemotherapy for gastric cancer. If the level of CEA drops to more than 50% or falls to the normal range and lasts for more than 4 weeks, it can be used as an effective indicator of treatment; (5) Its relationship with prognosis is still controversial, but most results believe that if Sustained increase after treatment indicates poor prognosis in patients with gastric cancer
However, there is no report on the combination of Cystatin SN and CEA markers in the detection of gastric cancer or gastrointestinal stromal tumors

Method used

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  • Combined application of cystatin SN and carcino-embryonic antigen
  • Combined application of cystatin SN and carcino-embryonic antigen
  • Combined application of cystatin SN and carcino-embryonic antigen

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] Example 1 Establishment of Cystatin SN serum detection reaction system and its optimization

[0028] 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; Concentrations are respectively 0pg / mL, 50pg / mL, 100pg / mL, 200pg / mL, 400pg / mL, 800pg / mL, 1600pg / mL Cystatin SN protein standard (the amino acid sequence of encoding Cystatin SN protein is as SEQ ID NO.1 shown) 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 ). The results showed that the linear range o...

Embodiment 2

[0035] Example 2 Establishment of CEA serum detection system and its optimization

[0036] Coat the ELISA plate with a mouse anti-human CEA monoclonal antibody at a concentration of 3 μg / mL, and the coating condition is to coat overnight at 4°C, wash the plate; then block it in 2% BSA at room temperature for 2 hours, wash plate; CEA protein standard (encoded amino acid sequence of CEA protein) (as shown in SEQ ID NO.2) and serum samples were added to the blocked plate, reacted at 37°C for 1 hour, and washed the plate; React 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 2 ). The results showed that the linear range of CEA was 39.0625pg / mL-1250pg / mL, the linear correlation coefficient r≥0.990 in the linear range, and the recovery rate was in the range of 90%-110%.

[0037] Detection system optimization: comparison of CEA...

Embodiment 3

[0043] Example 3 Cystatin SN-CEA enzyme-linked immunoassay kit

[0044] According to the establishment of Cystatin SN and CEA serum detection system in Example 1 and Example 2, the Cystatin SN-CEA enzyme-linked immunoassay kit was constructed, and the specific components are as shown in Table 1:

[0045] Table 1. Components of Cystatin SN-CEA ELISA Kit

[0046]

[0047]

[0048] Evaluation of Cystatin SN-CEA ELISA Kit: Use Cystatin SN-CEA ELISA Kit to test Cystatin SN Quality Control and CEA Quality Control 10 times at concentration levels of 160pg / mL and 80pg / mL respectively , the results showed that the coefficient of variation CV ≤ 10%; the same sample was tested with 3 batches of kits, and the results showed that the coefficient of variation between 3 batches of kits was CV ≤ 15%. The stability of the kit was also studied, and the results showed that it can be stored stably for 8 months under sealed conditions at 4°C, can be stored stably for 2 months under unsealed...

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Abstract

The invention discloses a combined application of a cystatin SN and a carcino-embryonic antigen (CEA), and particularly discloses the application of the cystatin SN and the CEA in preparation of a marker for detection of gastric cancer or gastrointestinal stromal tumor. The invention also discloses a capture agent of the gastric cancer or gastrointestinal stromal tumor marker and a kit containing the capture agent. The prepared kit has the advantages of good specificity, high sensitivity and the like, can be used for early diagnosis of gastric cancer or gastrointestinal stromal tumor, 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 combined application of cysteine ​​protease inhibitor SN and carcinoembryonic antigen. Background technique [0002] Gastric cancer is one of the malignant tumors that seriously threaten human health, and its mortality rate ranks second among all malignant tumors. At present, there are 934,000 new cases of gastric cancer every year in the world, of which nearly 400,000 are in China, and the morbidity and mortality are more than twice the world average. With the application of gastroscopy and modern imaging technology, the diagnosis rate of early gastric cancer has increased to about 10%, and the cure rate of early treatment has reached 95%. However, more than 90% of the hospitalized cases in China are in the middle and late stage. With medical treatment, the five-year survival rate is less than one-fifth. Improving the overall curative effect of gastric cancer largely depends ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68G01N33/574C07K16/38C07K16/18
CPCG01N33/57446G01N33/57484G01N2333/8139G01N2800/52G01N2800/54
Inventor 王弢秦勇
Owner SHANGHAI LIANGRUN BIOMEDICINE TECH CO LTD
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