Method for detecting discrepancy expressed protein spectrum in blood serum sample of diabetes and kidney disease patient

A technology for diabetic nephropathy and differential expression, which is applied in the fields of biology and medical testing, and can solve problems such as limited application

Inactive Publication Date: 2009-03-25
AFFILIATED HUSN HOSPITAL OF FUDAN UNIV
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  • Abstract
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  • Application Information

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Problems solved by technology

The application of SELDI-TOF-MS technology in diabetes research is extremely limited, and there is no report on large-scale research on the blood of patients with type 2 diabetic nephropathy and an analysis model

Method used

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  • Method for detecting discrepancy expressed protein spectrum in blood serum sample of diabetes and kidney disease patient
  • Method for detecting discrepancy expressed protein spectrum in blood serum sample of diabetes and kidney disease patient
  • Method for detecting discrepancy expressed protein spectrum in blood serum sample of diabetes and kidney disease patient

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Embodiment 1

[0018] Example 1: Establishment of Serum Protein Fingerprint and DN Decision Tree Model of Type 2 Diabetic Nephropathy

[0019] Take serum samples from patients with type 2 diabetes and normal people:

[0020] Patients diagnosed with type 2 diabetic nephropathy, patients diagnosed with type 2 diabetes without nephropathy, normal people, fasting venous blood 5ml (without jaundice, lipemia, hemolysis, etc.), after 3 hours at room temperature, centrifuge at 3000rpm at 4°C for 10min, take The upper serum was stored in a -80°C refrigerator.

[0021] Quantitative protein concentration by Bradford method: blank control: 1ml Coomassie brilliant blue solution + 100μl deionized water; prepare standard curve according to Table 1; measure sample concentration; protein sample plus corresponding binding buffer (WCX buffer or SAX buffer or IMAC buffer solution or H4 buffer), diluted to 1mg / ml and loaded onto the corresponding chip.

[0022] Table 1 Preparation of standard curve

[0023] ...

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Abstract

The invention belongs to the field of biology and medicine examination, and relates to a method for a protein spectrum of differential expression in a type 2 diabetes serum sample. The method comprises the following steps: adopting a surface-enhanced laser desorption/ionization time-of-flight mass-spectrometric technique to screen whole protein fingerprint of a diabetes chronic complication serum sample, using a data mining technique to screen biological labels in the fingerprint, establishing an analytic decision tree model of N serum protein components, and displaying that the type 2 diabetic nephropathy patient has the difference of the serum protein fingerprint as a result through blind verification and enlarged sample verification models. The method possibly provides reference information for further diagnosis and pathogenesis of diabetic nephropathy, provides research basis and new target for studying molecular mechanism formed by the diabetic nephropathy, and can further be used for guiding drug development and curative effect analysis.

Description

technical field [0001] The invention belongs to the field of biology and medical examination, and relates to a method for detecting body fluid markers of patients with diabetic nephropathy, in particular to a method for differentially expressed protein spectrum in serum specimens of type 2 diabetic nephropathy. Background technique [0002] The prior art shows that the main embodiment of biological function is protein, and protein expression profile can more truly reflect the functional mechanism of organisms than mRNA expression profile. Surface-enhanced laser desorption ionization time-of-flight mass spectrometry (SELDI-TOF-MS) is an ideal technical platform for current proteomics research, and it mainly consists of three parts: protein chip, mass spectrometer of flight and analysis software. The protein chip is the core part. According to the different properties of the chip, it can be divided into chemical surface chip and biological surface chip. The advent of protein ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N27/64G01N33/68
Inventor 胡仁明杨叶虹
Owner AFFILIATED HUSN HOSPITAL OF FUDAN UNIV
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