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Reagent kit for detecting 2-diabetes serum mark in Chinese people

A biologically active and molecular technology, applied in the field of detection kits for Chinese type 2 diabetes serum markers, can solve the problems of interfering with the isolation and identification of low-abundance proteins, and finding rare serum markers for diabetes

Inactive Publication Date: 2007-03-14
THE INST OF BASIC MEDICAL SCI OF CHINESE ACAD OF MEDICAL SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the presence of albumin in serum accounts for more than 80% of all proteins, which interferes with the isolation and identification of other low-abundance proteins
So far, the application of proteomics to find serum markers of diabetes is still very rare

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0019] Example 1 Two-dimensional electrophoresis and mass spectrometry analysis of differentially expressed proteins in the serum of diabetic patients and normal individuals

[0020] 1. Collection of samples from type 2 diabetic patients and normal controls in the Chinese Han population

[0021] The peripheral blood samples (10 ml) of the type 2 diabetes group and the normal group used in the present invention mainly come from the Department of Endocrinology, Peking Union Medical College Hospital. Diabetes cases can be sporadic or members of diabetic families (but at this time, only one patient is usually selected in each family, and more than one patient can be selected for members without any blood relationship in the same family); normal controls come from normal people, and their There are no diabetic patients (including type I diabetes) within three generations of the family, and the age and sex of the control group and the affected group are matched.

[0022] Diagnosis of...

Embodiment 2

[0039] Example 2 Detecting the expression level of the protein in type 2 diabetes patients and normal human serum by using enzyme-linked immunosorbent assay (ELISA)

[0040] 1) Serum coating Add serum diluted in PBS (1:1000 to 1:10000) to 100 μl / well of the enzyme-labeled well, and coat overnight at 4°C

[0041] 2) Standard protein coating Add standard protein serially diluted in PBS to 100 μl / well of the enzyme-labeled well and coat overnight at 4°C

[0042] 3) Blocking Dissolve skimmed milk powder in PBS to prepare 5% blocking solution, discard the coating solution, add blocking solution at 200 μl / well, 37°C, 2h. Pour off the blocking solution, and if not used immediately, dry it in an ultra-clean bench, seal it with tape, and store it at 4°C.

[0043] 4) Add the diluted anti-apolipoprotein A-I antibody (purified IgG, serum, ascites, etc.) to the enzyme-labeled well, 100 μl / well, overnight at 4°C, or 3-4 hours at 37°C.

[0044] 5) Pour off the primary antibody, wash the we...

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Abstract

The invention relates to the reagent box to detect serum of II-type diabetes. Specially, it applies the serum of apolipoprotein A-I and corresponding express-level in patient to modify the box. It also relates to the application in diagnosis and treatment of apolipoprotein A-I.

Description

technical field [0001] The present invention relates to a serum marker of type 2 diabetes, and the serum marker is apolipoprotein A-I, that is, subtype I of apolipoprotein A. The present invention also relates to a detection kit designed by utilizing the identified marker apolipoprotein A-I and the change of its expression level in patients. Further, the present invention also relates to the role of the serum marker in the diagnosis of diabetes, and as a target for drug design for diabetes treatment. Background technique [0002] As we all know, type 2 diabetes is a complex trait disease controlled by multiple genes, with obvious genetic predisposition. At present, it is believed that related protein molecules involved in insulin signal transduction pathway, glucose transport pathway, glycogen synthesis pathway, fatty acid absorption and synthesis, and adipocyte differentiation pathway may be related to the occurrence of diabetes. Research on the pathogenesis of diabetes, ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/53G01N33/68
Inventor 冯起平李云峰左瑾方福德
Owner THE INST OF BASIC MEDICAL SCI OF CHINESE ACAD OF MEDICAL SCI
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