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Combined application of cystatin S and carcinoembryonic antigen

A technology of cysteine ​​protease and carcinoembryonic antigen, which is applied in the field of medical detection, can solve the problems of poor prognosis, poor accuracy, undetected gastric cancer and gastrointestinal stromal tumor, etc., and achieve easy portability, convenient use, and repeatability Good results

Active Publication Date: 2014-07-09
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 detection of gastric cancer and gastrointestinal stromal tumors combined with Cystatin S and CEA markers

Method used

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  • Combined application of cystatin S and carcinoembryonic antigen
  • Combined application of cystatin S and carcinoembryonic antigen
  • Combined application of cystatin S and carcinoembryonic antigen

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] Example 1 Establishment of Cystatin S Serum Detection Reaction System and Its Optimization

[0029] 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; Concentrations are respectively 0pg / mL, 50pg / mL, 100pg / mL, 200pg / mL, 400pg / mL, 800pg / mL, 1600pg / mL Cystatin S protein standard substance (the amino acid sequence of coding Cystatin S 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 S polyclonal antibody labeled with HRP at a concentration of 0.5 μg / mL 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 ...

Embodiment 2

[0036] Example 2 Establishment of CEA serum detection system and its optimization

[0037] Coat the ELISA plate with a mouse anti-human CEA monoclonal antibody monoclonal antibody at a concentration of 3 μg / mL. The coating condition is to coat overnight at 4°C and wash the plate; then block in 2% BSA at room temperature for 2 hours, wash the plate; the concentration of 0pg / mL, 39.0625pg / mL, 78.125pg / mL, 156.25pg / mL, 312.5pg / mL, 625pg / mL, 1250pg / mL) CEA protein standard (coding CEA protein The amino acid sequence is 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 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 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...

Embodiment 3

[0044] Example 3 Cystatin S-CEA enzyme-linked immunoassay kit

[0045] According to the establishment of Cystatin S and CEA serum detection system in Example 1 and Example 2, the Cystatin S-CEA enzyme-linked immunoassay kit was constructed, and the specific components are as shown in Table 1:

[0046] Table 1. Cystatin S-CEA enzyme-linked immunoassay kit components

[0047]

[0048]

[0049] Evaluate the Cystatin S-CEA ELISA Kit: Use the Cystatin S-CEA ELISA Kit to repeat the Cystatin S positive quality control and positive CEA quality control at concentrations of 160pg / mL and 80pg / mL, respectively Tested 10 times, 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 inter-assay coefficient of variation of the 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...

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Abstract

The invention discloses combined application of cystatin S and carcinoembryonic antigen (CEA), and specifically discloses application of cystatin S and CEA to prepare a marker for diagnosing and indicating stomach cancer or gastrointestinal stromal tumor. The invention also discloses a trapping agent of the marker of stomach cancer or gastrointestinal stromal tumor and a kit containing the trapping agent. The kit has the advantages of being good in specificity, high in sensitivity and the like, is applicable to early diagnosis on stomach cancer or gastrointestinal stromal tumor, 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 combined application of cysteine ​​protease inhibitor S 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 o...

Claims

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Application Information

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IPC IPC(8): G01N33/68G01N33/574C07K16/38C07K16/18
CPCC07K16/3007C07K16/38G01N33/57446G01N33/57473G01N2333/8139G01N2800/52G01N2800/54
Inventor 王弢秦勇渠香云
Owner SHANGHAI LIANGRUN BIOMEDICINE TECH CO LTD
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