Method for detecting type 2 diabetic salivary protein marker

A type 2 diabetes, salivary protein technology, applied in measurement devices, instruments, scientific instruments, etc., can solve problems such as insufficient insulin formation and elevated blood sugar

Inactive Publication Date: 2018-06-15
SHENZHEN ELDERLY MEDICAL RES INST +1
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Problems solved by technology

With the advancement of science, people's research on type 2 diabetes has gradually shifted from gene to gene expression protein research. For example, it was found that the content of prohormone convertase (Prohormone convertase, PC) in type 2 diabetes was significantly higher than that of normal people. This enzyme is an endoprotease that can process the precursor of neuroendocrine hormones, and can act on pancreatic β cells to remove part of the peptide from the secreted proinsulin to form insulin. Can lead to insufficient formation of insulin, which can lead to elevated blood sugar

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  • Method for detecting type 2 diabetic salivary protein marker
  • Method for detecting type 2 diabetic salivary protein marker
  • Method for detecting type 2 diabetic salivary protein marker

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Embodiment Construction

[0090] The technical solution of this patent will be further described in detail below in conjunction with specific embodiments.

[0091] see Figure 1-4 , a kind of detection method of type 2 diabetes salivary protein marker, concrete steps are as follows:

[0092] (1) Sample protein extraction: Add 5% lysis buffer to the sample, vortex mix, ultrasonicate for 60s, amplitude 22%, extract at room temperature for 30min; centrifuge at 15000r / min, 4°C for 20min, take out the supernatant; Freeze at -80°C after aliquoting;

[0093] (2) Protein quantification: use the Bradford method to measure the extracted protein concentration. First, dilute the sample with lysis buffer to make the final concentration fall within the range of the calibration curve. Take 10 μl and 300 μl of the diluted sample and standard respectively. The protein quantitative dye was reacted in the dark for 20 minutes, and the absorbance value of the standard substance and the sample at 595 nm was measured simul...

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Abstract

The invention discloses a method for detecting a type 2 diabetic salivary protein marker. The method comprises extracting a sample protein, acquiring the concentration of the extracted protein througha Bradford method, carrying out protein proteolysis, carrying out iTRAQ labeling, carrying out vacuum refrigeration and centrifugal drying, carrying out reversed-phase chromatography separation at high pH, carrying out ABI-5600-based protein analysis and mass spectrometry data analysis, in database detection, if the sample belongs to the sequenced organism, directly selecting the database of thesequenced organism, if the sample does not belong to the sequenced organism, selecting a database of a macroproteome with the largest correlation with the tested sample, and carrying out iTRAQ mass spectrometric analysis through the ABI-5600 mass spectrometer. The method can fast and reliably find a specific protein marker of diabetes through a proteome technical scheme and build a non-invasive, simple, rapid and practical detection and assessment method for early diagnosis, postoperative recurrence, metastasis and prognosis observation of diabetes.

Description

technical field [0001] The invention relates to a method for detecting biomarkers, in particular to a method for detecting salivary protein markers of type 2 diabetes. Background technique [0002] Type 2 diabetes (Type 2diabetes mellitus, T2DM) is a common clinical metabolic disorder in the elderly. The biochemical examination mainly shows the increase of fasting blood glucose and (or) postprandial blood glucose. Typical symptoms may be accompanied by dry mouth and polydipsia. , eating too much and easily hungry, increased urine output, and weight loss. In the later stage of the disease, patients often have atherosclerotic lesions in the heart, brain, kidney and other tissues and organs, resulting in narrowing of the vascular lumen, resulting in a series of heart, Clinical manifestations of insufficient blood supply, metabolic disorders or even failure of brain, kidney and other tissues and organs. In critically ill patients, due to serious abnormalities in the metabolism ...

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Application Information

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IPC IPC(8): G01N30/89
CPCG01N30/89
Inventor 吴正治黄飞娟孙珂焕曹美群丁峰范大华姚永超
Owner SHENZHEN ELDERLY MEDICAL RES INST
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