Detection method of diabetes B kidney deficiency salivary protein markers

A technology for type 2 diabetes and a detection method, which is applied in the field of detection of saliva protein markers of type 2 diabetes with kidney deficiency syndrome, can solve the problems affecting the feasibility and operability of traditional Chinese medicine for the treatment of type 2 diabetes, the variety of syndrome differentiation and typing methods of type 2 diabetes, and the lack of Quantitative indicators, etc.

Inactive Publication Date: 2018-05-01
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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  • Detection method of diabetes B kidney deficiency salivary protein markers
  • Detection method of diabetes B kidney deficiency salivary protein markers
  • Detection method of diabetes B kidney deficiency salivary protein markers

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Embodiment Construction

[0265] The technical solution of this patent will be further described in detail below in conjunction with specific embodiments.

[0266] see Figure 1-4 , a method for detecting salivary protein markers of type 2 diabetes with kidney deficiency syndrome, the specific steps are as follows:

[0267] (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;

[0268] (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 detection method of diabetes B kidney deficiency salivary protein markers. The method comprises the following concrete steps of extracting sample protein; determining the extracted protein concentration by a Bradford method; performing proteolysis, iTRAQ marking, vacuum freeze centrifugation drying and reversed-phase chromatography separation under high pH conditions; performing protein analysis on ABI-5600; performing mass spectrometric data analysis; during database selection, if organisms are sequenced organisms, directly selecting the species databases; if the organisms are non-sequenced organisms, selecting main category proteome database most relevant to the tested sample; using an ABI-5600 type mass spectrograph to perform iTRAQ mass spectrograph analysis.The specific protein marker in the diabetes B kidney deficiency can be fast and reliably discovered by a proteomic technological scheme. A noninvasive, simple, convenient, fast and practical detectionand evaluation measure is built for the early diagnosis and treatment, postoperative recurrence, transfer and prognosis observation of the diabetes B kidney deficiency.

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 kidney deficiency syndrome. Background technique [0002] Type 2 diabetes 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 frequent Drinking, eating too much, easy hunger, 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, which leads to a series of heart disease. The clinical manifestations of insufficient blood supply, metabolic disorders and even failure of tissues and organs such as brain, kidney, etc. In critically ill patients, due ...

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68
CPCG01N33/6848G01N33/6893G01N2800/042
Inventor 吴正治孙珂焕黄飞娟曹美群丁峰范大华姚永超
Owner SHENZHEN ELDERLY MEDICAL RES INST
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