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CEA negative colorectal cancer detection and prognosis judgement mass spectrum reagent kit and method

A colorectal cancer and kit technology, applied in the field of protein detection, can solve the problems of difficult to obtain reproducibility of pathological evidence, lack of early monitoring methods, and no obvious clinical manifestations of tumors.

Inactive Publication Date: 2008-08-13
许洋
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the clinical evaluation of the prognosis of malignant tumors mainly relies on clinical manifestations, pathology and traditional tumor markers. However, there are often no obvious clinical manifestations when tumors recur or metastasize in the early stage, and pathological evidence is difficult to obtain and has poor reproducibility.
The traditional tumor marker carcinoembryonic antigen (CEA) is of great value in the prognosis assessment of colorectal cancer, but it is also found in clinical work that a considerable proportion of colorectal cancer patients cannot be detected due to the negative expression of carcinoembryonic antigen. The prognosis assessment of colorectal cancer patients with negative expression lacks effective early monitoring methods
However, when we performed protein mass spectrometry analysis, we found that there was no standardized kit for mass spectrometry of clinical CEA-negative colorectal cancer

Method used

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  • CEA negative colorectal cancer detection and prognosis judgement mass spectrum reagent kit and method
  • CEA negative colorectal cancer detection and prognosis judgement mass spectrum reagent kit and method
  • CEA negative colorectal cancer detection and prognosis judgement mass spectrum reagent kit and method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0063] Example 1 Identification of normal and CEA-negative colorectal cancer patients and preparation of mass spectrometry kits

[0064] (1) Experimental method

[0065]60 patients with CEA-negative colorectal cancer (CEA≤5μg / L) and 68 patients with CEA-positive colorectal cancer (CEA>5μg / L), all of them were adenocarcinoma, with an average age of 61.5 years; only 20 patients with advanced colorectal cancer No lymph node metastasis was seen in 1 case, and the rest were associated with lymph node metastasis; 32 cases had organ metastasis: 16 cases of liver metastasis, 12 cases of lung metastasis, and 4 cases of ovarian metastasis. Fifteen patients with colorectal benign tubular adenoma confirmed by colonoscopy. 60 healthy subjects, with an average age of 59.5 years, were from a population with normal liver function and renal function. Collect 1mL of venous blood from the subject on an empty stomach, immediately after collection, let it stand in a refrigerator at 4°C for 2 hou...

Embodiment 2

[0082] The double-blind test of embodiment 2 kit

[0083] Several characteristic protein peaks were screened out from Example 1, and the test model composed of 3 differential peaks of 3398.3, 5477.1, and 8453.9 ± 15Da had a 100% sensitivity to the blind screening test of carcinoembryonic antigen-negative colorectal cancer patients and healthy people , specificity 83% (Table 2); that is, when 3398.3Da is less than or equal to 7.83; or when 3398.3Da is greater than 7.83 and 5477.1Da is greater than 1.54 and 8453.9Da is greater than 1.18, it is a colorectal cancer patient with negative expression of carcinoembryonic antigen, otherwise it is a healthy population ( image 3 , Table 1, 2).

[0084] Table 2 Double-blind test of kits

[0085] double blind test

Carcinoembryonic antigen-negative colorectal cancer (60)

normal(60)

carcinoembryonic antigen negative colorectal cancer

60

10

normal

0

50

[0086] Sensitivity 100...

Embodiment 3

[0088] Example 3 Screening of colorectal cancer metastasis-associated polypeptides

[0089] Patients with colorectal cancer were grouped according to organ metastasis, and then compared between groups, among which 4 polypeptides and proteins were significantly increased in the organ metastasis group (Table 3). The colorectal cancer patients were grouped according to the lymph node metastasis group, and it was found that the two polypeptides and proteins were significantly increased in the lymph node metastasis group (Table 4). The 11683.3Da peak of the same representative patient shows that the sensitivity of WCX magnetic beads is higher than that of WCX chips ( Figure 4 ).

[0090] Table 3 Differences in the abundance of polypeptide proteins between colorectal cancer organ metastasis group and non-organ metastasis group (P<0.05, x±sd)

[0091]

[0092] Table 4 Differences in polypeptide protein abundance between colorectal cancer lymph node metastasis group and non-lymp...

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Abstract

The present invention relates to a magnetic beads support substrate method to capture colorectal cancer proteome in biology sample using magnetic separator separating magnetic beads and sample without centrifugation sample, and protein fingerprint method analyzed by mass spectrometry. The present invention is suitable for detecting person and CEA negative colorectal cancer and reagent kits of prognosis estimation protein fingerprint or mass spectrometry polypeptide atlas. The method of present invention is accurate, convenient and rapid.

Description

technical field [0001] The invention relates to a kit for a new method for analyzing proteins in colorectal cancer biological samples. A biomarker captured by a protein-binding matrix and quantitatively controlled mass spectrometry for the detection of CEA-negative colorectal cancer biomarkers. The invention mentioned here relates to the field of protein detection and is a new non-invasive in vitro mass spectrometry detection method. The present invention can be applied to the detection method or kit of the CEA negative colorectal cancer biomarker combination in the body fluid that has been separated from the human body. Background technique [0002] With the implementation and completion of the Human Genome Project, scientists proposed the concept of the post-genome (post-genome) project, and the focus of research shifted to functional genomics, and the main substance of biological function is protein. In 1994, Wilkins and Williams of Macquarie University in Australia fir...

Claims

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Application Information

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IPC IPC(8): G01N33/68G01N30/02
Inventor 许洋
Owner 许洋
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