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Method for detecting chronic nephritis biomarkers based on salivary proteomics

A technology of proteomics and biomarkers, applied in the field of detection of chronic nephritis biomarkers based on saliva proteomics, can solve the problems of no practical significance, kidney damage, etc.

Inactive Publication Date: 2018-04-03
SHENZHEN ELDERLY MEDICAL RES INST +1
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Problems solved by technology

Since protein in urine is closely related to kidney function, changes in protein in urine indicate that the kidney has been damaged. Therefore, it is meaningless to look for biomarkers of nephritis in urine

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  • Method for detecting chronic nephritis biomarkers based on salivary proteomics
  • Method for detecting chronic nephritis biomarkers based on salivary proteomics
  • Method for detecting chronic nephritis biomarkers based on salivary proteomics

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[0244] The technical solution of this patent will be further described in detail below in conjunction with specific embodiments.

[0245] see figure 1 , a method for detecting chronic nephritis biomarkers based on salivary proteomics, the specific steps are as follows:

[0246] (1) Sample protein extraction: add 5% lysis buffer to the sample, vortex mix, sonicate 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;

[0247] (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 measure...

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Abstract

The invention discloses a method for detecting chronic nephritis biomarkers based on salivary proteomics. The method comprises the following specific steps: sample protein extraction; determination ofthe concentration of extracted protein by using a Brad ford method; proteolysis; iTRAQ marking; vacuum refrigerated centrifugation drying; reversed phase chromatography under a high pH condition; protein analysis by using ABI-5600; mass spectral data analysis, namely when selecting a database, in case of an sequenced organism, a database for this species is directly selected, and in case of a non-sequenced organism, a database for a large-range proteome most relevant to a sample to be tested is selected; iTRAQ mass spectrometric analysis by using an ABI-5600 mass spectrometer. The method canbe used for finding out specific protein markers in chronic nephritis by using quick and reliable proteomics technology scheme, so as to establish a non-invasive, easy, convenient, fast, and practicaldetection assessment for early diagnosis and treatment, postoperative recurrence, metastasis and prognosis observation of the chronic nephritis.

Description

technical field [0001] The present invention relates to a method for detecting biomarkers, in particular to a method for detecting biomarkers of chronic nephritis based on salivary proteomics. Background technique [0002] Nephritis is inflammation of the kidney, which may include glomeruli, tubulointerstitium, or glomerular and peritubular tissue, with the participation of immune-mediated, inflammatory mediators (such as complement, cytokines, reactive oxygen species, etc.), and finally A group of kidney diseases that lead to inflammatory changes in the renal proper tissues and various degrees of renal dysfunction can be caused by a variety of etiologies, and non-immune and non-inflammatory mechanisms are also involved in the chronic process. How to diagnose renal disease and carry out early intervention is an urgent problem to be solved clinically. The detection of biomarkers can realize the early diagnosis of kidney disease, the assessment of the patient's prognosis and ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68
CPCG01N33/6848G01N2800/347G01N2800/60
Inventor 吴正治姚永超曹美群孙珂焕丁峰黄飞娟范大华
Owner SHENZHEN ELDERLY MEDICAL RES INST
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