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ELISA (enzyme linked immunosorbent assay) kit for detecting endothelial cell specific molecule-1 (ESM-1) of tumour marker

An endothelial cell-specific and kit technology is applied in the field of kits for detecting endothelial cell-specific molecule-1, which can solve the problems of gastric cancer specificity and low specificity, and achieve the effect of improving the early diagnosis rate.

Inactive Publication Date: 2011-09-07
BEIJING CANCER HOSPITAL PEKING UNIV CANCER HOSPITAL
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Tumor markers CA19-9, CA72-4 and CA242 also have the same problem, and the specificity is not high
Therefore, there is no tumor marker that is completely specific for gastric cancer

Method used

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  • ELISA (enzyme linked immunosorbent assay) kit for detecting endothelial cell specific molecule-1 (ESM-1) of tumour marker
  • ELISA (enzyme linked immunosorbent assay) kit for detecting endothelial cell specific molecule-1 (ESM-1) of tumour marker
  • ELISA (enzyme linked immunosorbent assay) kit for detecting endothelial cell specific molecule-1 (ESM-1) of tumour marker

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] Example 1 The level of ESM-1 nucleic acid in tumor tissue is significantly up-regulated, and the expression of ESM-1 mRNA and protein in gastric cancer tissue is up-regulated:

[0030]In the early stage, we used high-density oligonucleotide chip technology to conduct comprehensive and large-scale research on gastric cancer-related genes, using 42,292 long oligonucleotides (24,000 are reference sequences in the gene database (RefSeqdatabase release 17), The other 24,000 items are EST and Unigene sequence (Unigene build188)) probe chips were studied on the first batch of 46 cases of gastric cancer and 20 cases of non-tumor gastric mucosa, and the differential expression between gastric cancer and adjacent normal tissues and the biological characteristics of gastric cancer were screened out gene. These genes can be used to classify tumors, explore the possible etiology of tumors, predict the prognosis of tumor patients and determine the relevant targets of tumor molecular ...

Embodiment 2

[0071] Example 2 Analysis of the relationship between ESM-1 expression and clinicopathological parameters

[0072] The relationship between the expression of ESM-1 in gastric cancer tissues and the clinicopathological features is shown in Table 7. Chi-square test analysis showed that the positive expression of ESM-1 was more likely to occur in patients with vascular invasion, distant metastasis and Borrmann type IV. The incidence of vascular invasion in patients with positive expression of ESM-1 was 67% (61 / 91), which was significantly higher than 45.3% (29 / 64) in the non-expression group. In addition, the incidence of distant metastasis was also different between the two groups (17.4% vs. 7.5%) (P0.05).

[0073] Table 7 Relationship between ESM-1 expression and clinicopathological parameters (159 patients with gastric cancer)

[0074]

[0075]

[0076] Kaplan-Meier survival analysis was used to evaluate the relationship between the expression of ESM-1 and the clinical...

Embodiment 3

[0082] Example 3: Determination of antibodies and detection conditions for detecting ESM-1

[0083] ESM-1 human recombinant protein (1810EC; R&D Systems, USA) was used as a standard to make a standard curve, and a sensitive and specific paired coating antibody and detection antibody were selected to determine the detection conditions by square array titration.

[0084] Determine the working concentration of the antibody: Dilute the coated antibody with coating solution to four concentrations of 4 μg / mL, 2 μg / mL, 1.3 μg / mL, and 1 μg / mL, and coat the 96-well plate (96-well microtiter plate, Corning company, the U.S.), each concentration was coated with three lines (3 wells per line), and a strong positive antigen was added to the first, second, and third lines of each concentration coating (the concentration of ESM-1 human recombinant protein was 10000pg / ml), weak positive antigen (ESM-1 human recombinant protein concentration is 2000pg / ml) and negative control (normal human se...

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Abstract

The invention provides an ELISA (enzyme linked immunosorbent assay) kit for detecting endothelial cell specific molecule-1 (ESM-1) of a tumour marker, wherein the kit comprises (1) a coated antibody, wherein the coated antibody is a mouse anti-human ESM-1 antibody; (2) a detection antibody, wherein the detection antibody is a biotinylated goat anti-human Endocan antibody; (3) an enzyme linked antibody, wherein the enzyme liked antibody is a rabbit anti-goat IgG (immunoglobulin G) marked by a horseradish peroxidase; (4) a human Endocan standard substance; and (5) a colour development substrate of the enzyme linked antibody. By the kit provided by the invention, the early diagnosis rate of gastric cancer can be improved under a relatively small wound; the biomarker is an independent prognostic factor and is relevant to vessel invasion, distant metastasis and general classification of gastric cancer, so that prognosis can be predicted, states of the illness can be monitored, and treatments can be guided.

Description

technical field [0001] The invention relates to a kit for detecting tumor markers, in particular to a kit for detecting endothelial cell specific molecule-1 (ESM-1). Background technique [0002] Gastric cancer is one of the most common malignant tumors that threaten human health, and is the second most lethal tumor. Surgical resection is currently the only possible cure. However, most patients are already at an advanced stage when they are discovered and cannot be treated surgically. Therefore, timely and accurate diagnosis is the key to saving patients' lives. [0003] Existing detection methods for gastric cancer include tissue section immunohistochemistry. However, the immunohistochemical technique has many steps, high requirements on experimenters and experimental conditions, and there are many false positive and false negative staining. Therefore, the results of immunohistochemistry are not stable enough (see references 6, 7, 8, 9, 10). The most unfavorable thing is...

Claims

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Application Information

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IPC IPC(8): G01N33/543G01N33/53
Inventor 季加孚朱昱冰刘妮张连海王晓红邢晓芳杜红胡颖
Owner BEIJING CANCER HOSPITAL PEKING UNIV CANCER HOSPITAL
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