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Testing method of type-II diabetes mellitus spleen-deficiency syndrome sialoprotein marker

A type 2 diabetes and detection method technology, applied in the direction of measuring devices, instruments, scientific instruments, etc., can solve the problems affecting the feasibility and operability of traditional Chinese medicine treatment of type 2 diabetes, the variety of syndrome differentiation and typing methods of type 2 diabetes, and the lack of quantitative indicators And other issues

Inactive Publication Date: 2018-06-12
SHENZHEN ELDERLY MEDICAL RES INST +1
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Problems solved by technology

However, in real life, the diagnosis and treatment of type 2 diabetes with traditional Chinese medicine has not been well valued and widely used. The main reason is that there are too many methods of syndrome differentiation and typing of type 2 diabetes, and the lack of reliable objective quantification indicators, thus affecting the feasibility and specific operability of traditional Chinese medicine in treating type 2 diabetes to a considerable extent

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  • Testing method of type-II diabetes mellitus spleen-deficiency syndrome sialoprotein marker
  • Testing method of type-II diabetes mellitus spleen-deficiency syndrome sialoprotein marker
  • Testing method of type-II diabetes mellitus spleen-deficiency syndrome sialoprotein marker

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[0141] The technical solution of this patent will be further described in detail below in conjunction with specific embodiments.

[0142] see Figure 1-4 , a method for detecting salivary protein markers of type 2 diabetes with spleen deficiency syndrome, the specific steps are as follows:

[0143] (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;

[0144] (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 sa...

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Abstract

The invention discloses a testing method of a type-II diabetes mellitus spleen-deficiency syndrome sialoprotein marker. The testing method comprises the following specific steps: extracting sample protein; measuring the concentration of the extracted protein by a Brad ford method; performing proteolysis; performing iTRAQ (isobaric tags for relative and absolute quantification) labeling; performingvacuum freezing, and centrifugal drying; performing reversed phase chromatographic separation in a high-pH condition; performing protein analysis by using ABI-5600; performing mass spectrometric dataanalysis: when selecting a database, for the sequenced organism, directly selecting the database of the species thereof; for the non-sequenced organism, selecting the database of large-class proteomemost related to the detected sample; performing mass spectrometry on iTRAQ by using an ABI-5600 type mass spectrometer. The method disclosed by the invention can discover a specific protein marker inthe diabetes mellitus spleen-deficiency syndrome by using a rapid and reliable proteomics technical scheme, and creates a non-invasive, simple, quick and practical detection and evaluation means forearly-stage diagnosis and treatment, postoperative recurrence, metastasis and prognosis observation of the diabetes mellitus spleen-deficiency syndrome.

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 mellitus with spleen deficiency syndrome. 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...

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Application Information

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IPC IPC(8): G01N30/89
CPCG01N30/89
Inventor 吴正治孙珂焕姚永超黄飞娟曹美群丁峰范大华
Owner SHENZHEN ELDERLY MEDICAL RES INST
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