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Serum protein marker group for diagnosing MODY (maturity-onset-diabetes of the young) and application thereof

A serum protein and diabetes technology, applied in the field of diabetes diagnostic protein markers, can solve the problems of diagnosis deviation and poor treatment effect, and achieve reliable and accurate diagnosis results

Inactive Publication Date: 2017-05-10
XINJIANG MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Diagnostic bias is one of the main causes of poor treatment outcomes at present

Method used

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  • Serum protein marker group for diagnosing MODY (maturity-onset-diabetes of the young) and application thereof
  • Serum protein marker group for diagnosing MODY (maturity-onset-diabetes of the young) and application thereof
  • Serum protein marker group for diagnosing MODY (maturity-onset-diabetes of the young) and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] Differentially expressed proteins in patients with MODY and healthy controls detected by iTRAQ technology

[0034] 1. Test samples:

[0035] A four-generation surviving Uyghur family with premature diabetes. Divided into 4 cases of case and 4 cases of normal control serum. Collect 2mL of whole blood on an empty stomach in the morning, let it stand at 4°C for 1-2h until the blood coagulates and precipitate the serum, centrifuge at 3000g for 10min, collect the supernatant, aliquot it on ice and store it at -80°C for later use.

[0036] 2. Detection method:

[0037] (1) Use the Proteominer kit to remove high-abundance proteins in serum: add DTT with a final concentration of 10mM to the enriched protein sample, and bathe in water at 56°C for 1h; after cooling to room temperature, add IAM with a final concentration of 55mM, and place in a dark room for 45min Add 1mL of cold acetone to precipitate overnight, centrifuge at 25000rpm*4°C for 15min and discard the supernatant;...

Embodiment 2

[0047] Validation of Mass Spectrometry MRM in MODY Family

[0048] 1. Test samples:

[0049] MODY patients, 4 cases of normal control serum; all from the family

[0050] 2. Detection method:

[0051] The establishment of the experimental method 1) Select the MODY-related target protein suitable for MRM detection and analysis; 2) Evaluate the quality of the extracted protein; 3) Select the parent-child ion pair suitable for MRM detection; 4) Analyze the mass spectrum based on the analysis software Skyline 5) Perform quality assessment and analysis on the obtained detection data. Sample Analysis:

[0052] (1) Serum proteolysis: 12.5ug of each sample was taken, combined into two tubes of MODY case group and healthy control group, and 01ug / ul of BSA was added to each tube as an internal reference. Use an ultrafiltration tube, take the protein solution, centrifuge for 20min, remove the bottom solution, add 100ul TEAB, centrifuge as above, repeat the step 3 times, replace a new ...

Embodiment 3

[0061] 1. Test samples:

[0062] Type 1 diabetes patients, type 2 diabetes patients, 10 cases of healthy control serum;

[0063] 2. Detection method:

[0064] The establishment of the experimental method 1) MRM detection and analysis of the selected parent-child ion pairs in the expanded 30 samples; 2) Based on the analysis software Skyline, the mass spectrometer scanning parameters-collision energy were optimized; 4) The obtained detection Data quality assessment and analysis.

[0065] Sample analysis:

[0066] (1) Serum proteolysis: 200ug was taken from each sample, and 0.1ug / ul BSA was added to each tube as an internal reference. Add DTT with a final concentration of 10mM to the taken original serum sample, and put it in a water bath at 56°C for 1h; after cooling to room temperature, add IAM with a final concentration of 55mM, and place it in a dark room for 45min; add TEAB with a final concentration of 100Mm, according to the protein:enzyme=40: Add Trypsin enzyme at a ...

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Abstract

The invention relates to a serum protein marker group for diagnosing MODY (maturity-onset-diabetes of the young) and application thereof. The serum protein marker group comprises five proteins: apolipoprotein C-IV (APOC4), apolipoprotein a (LPA), a complement component C6 (C6), a blood coagulation factor v (F5) and thyroid binding globulin (SERPINA7). ITRAQ (isobaric tags for relative and absolute quantitation) are used to combine with an MALDI-TOF / MS technology to detect; mass spectrum detection shows that expression levels of the serum apolipoprotein C-IV, the apolipoprotein a, the complement component C6 and the blood coagulation factor v protein in serum of a MODY patient are obviously improved, and an expression level of the thyroid binding globulin in the serum of the MODY patient is obviously reduced. Mass spectrum MRM (multiple-reaction-monitoring) also verifies that the five proteins are abnormal in expression in serums of the MODY patient, type-1 and type-2 diabetics and a healthy control group, and are specific proteins.

Description

technical field [0001] The invention relates to the field of diabetes diagnostic protein markers, in particular to a serum protein marker group for diagnosing MODY of early onset diabetes and its application. Background technique [0002] Maturity-onset-diabetes of the young (MODY) is a monogenic dominant genetic disease. It was previously considered a special subtype of T2DM, but now it is classified as a special type of diabetes. The diagnosis of MODY should meet the following conditions: ①Onset before the age of 25; ②At least 2 patients in the family; ③At least three generations of autosomal dominant inheritance; ④Oral hypoglycemic drugs have been effective for at least 5 years or have normal plasma C-peptide levels. [0003] With the development of technologies such as genetic linkage analysis, candidate gene cloning, association analysis, and sequencing, by early 2013, 13 relatively definite genes related to the onset of MODY had been discovered, which are (according to...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68C07K14/775C07K14/47C12N9/64
CPCC07K14/47C07K14/472C07K14/775C12N9/6424G01N33/6893G01N2800/042
Inventor 帕它木·莫合买提先锋热沙来提·阿不都瓦衣特伊力哈木江·依马木木哈达斯·吐尔逊依明热比亚·努力伊再提古丽·木提拉祖力卡提阿衣·阿布都拉阿瓦古丽·托合提艾扎提古丽·卡的尔
Owner XINJIANG MEDICAL UNIV
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