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Biomarkers for diagnosing mucopolysaccharidosis type II (MPS II) and application thereof

A mucopolysaccharidosis and protein marker technology, applied in the field of biomarkers for the diagnosis of mucopolysaccharidosis type II, can solve the problems that the pathogenic mechanism has not been fully elucidated and hinders the exploration of disease treatment methods

Inactive Publication Date: 2019-05-21
GENERAL HOSPITAL OF PLA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At the same time, the pathogenic mechanism of the disease has not been fully elucidated, hindering the exploration of disease treatment methods

Method used

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  • Biomarkers for diagnosing mucopolysaccharidosis type II (MPS II) and application thereof
  • Biomarkers for diagnosing mucopolysaccharidosis type II (MPS II) and application thereof
  • Biomarkers for diagnosing mucopolysaccharidosis type II (MPS II) and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] Example 1. Collect urine samples from MPSⅡ patients and health indicators, and identify differentially expressed proteins by 2D-PAGE combined with MALDI-TOF / TOF.

[0024] 1. Precipitate protein by acetone method:

[0025] (1) Add 3-5 times the volume of acetone-containing solution (containing 0.07% β-mercaptoethanol) to the sample, and leave it at -20°C for 2 hours or overnight;

[0026] (2) Centrifuge at 12000 g for 5 min at low temperature, wash with 80% acetone (containing 0.07% β-mercaptoethanol) for 3 times, add appropriate amount of lysate, sonicate at low temperature for 5 min, and let stand at 4°C for more than 2 hr.

[0027] 2. Quantification of protein samples by Bradford method:

[0028] Bradford working solution formula (500ml): (1) 425ml double distilled water, (2) 15ml 95% ethanol, (3) 30ml 85% phosphoric acid, (4) 30ml Bradford stock solution (100mg Coomassie brilliant blue + 50ml 95% ethanol + 100ml 85% orthophosphoric acid, add double distilled water ...

Embodiment 2

[0065] Embodiment 2, ELISA detects the expression difference of target protein in pathological sample and normal sample

[0066] Select three proteins with large expression differences, alpha1-antitrypsin (AAT), Gm2activator (GM2A) and lipocalin-type prostaglandin D synthase (L-PGDS), to detect the expression difference of the target protein in pathological samples and normal samples

[0067] 1. AAT content detection

[0068] (1) Take the α1-AT standard substance (400ng / ml), and prepare solutions with concentrations of 200, 100, 50, 25, 12.5, and 6.25ng / ml with the standard dilution solution to establish the standard curve.

[0069] (2) Take out the urine sample from the refrigerator at -80°C, centrifuge at 12 000r / min at 4°C for 3min, and take the supernatant.

[0070] (3) Set up 7 standard wells, add 100 μl of standard products of different concentrations in sequence, and add 100 μl of ultrapure water to the blank well.

[0071] (4) Add 100 μl of the 10-fold diluted sample...

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Abstract

The invention relates to a group of biomarkers for diagnosing mucopolysaccharidosis type II (MPS II) and application thereof. The biomarkers are alpha 1-antitrypsin (AAT) in urine, Gm2 activator (GM2A) and lipocalin-Type prostaglandin D synthase (L-PGDS).

Description

technical field [0001] The invention belongs to the technical field of biomedicine, and in particular relates to a biomarker for diagnosing mucopolysaccharidosis type II and its application. Background technique [0002] Mucopolysaccharidosis type II (MPS II), also known as Hunter syndrome, is the most common type of MPS reported in my country. It is caused by iduronate-2-sulfatase (IDS) in lysosomes. ) deficiency, which leads to the accumulation of incompletely decomposed dermatin sulfate (dermatin sulfate, DS) and heparan sulfate (heparan sulfate, HS) in various tissues and organs of the body, and is a metabolic disease with X recessive inheritance. The clinical manifestations of the disease are general bone dysplasia, rough face, hepatosplenomegaly and so on. Most children with severe disease develop symptoms at 18-36 months, die at puberty, and have a very poor prognosis. Early diagnosis, especially for infants, will cause irreversible changes in the patient's developme...

Claims

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Application Information

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IPC IPC(8): G01N33/68
Inventor 王成彬段晋燕苑晓舟孟岩
Owner GENERAL HOSPITAL OF PLA
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