Application of protein combined marker in preparation of children asthma diagnostic reagent

A childhood asthma and protein technology, applied in disease diagnosis, biological testing, biomaterial analysis, etc., can solve the problems of poor treatment effect of children's cough variant asthma, achieve simple and easy sampling detection, realize high-throughput detection, Prepare simple effects

Active Publication Date: 2020-08-07
HANGZHOU NORMAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In terms of treatment effect, some children with asthma are not sensitive to corticosteroids, and antibiotic therapy is not effective in treating cough variant asthma in children, and

Method used

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  • Application of protein combined marker in preparation of children asthma diagnostic reagent
  • Application of protein combined marker in preparation of children asthma diagnostic reagent
  • Application of protein combined marker in preparation of children asthma diagnostic reagent

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] Collection of serum samples from children with asthma and healthy children:

[0035] Serum samples were collected from 135 childhood asthma patients and 55 healthy children. Take 3 mL of fasting peripheral venous whole blood, place it vertically at 4°C for 1-2 hours, centrifuge at 4000g for 10 minutes, and then collect the serum.

Embodiment 2

[0037] Isotope labeling relative and absolute quantitative labeling combined with two-dimensional liquid chromatography tandem mass spectrometry to screen differential proteins in children with asthma:

[0038] Serum samples from 15 children with asthma and 15 healthy children were randomly selected and mixed into a sample pool. Removal of 14 high-abundance proteins, including albumin, haptoglobin, IgG, complement C3, transferrin, antitrypsin, from a pool of serum samples from children with asthma and healthy controls using a Human 14 Multiple Affinity Removal System column , IgA, fibrinogen, transthyretin, α2-macroglobulin, IgM, apolipoprotein AI, α1-acid glycoprotein, apolipoprotein AII. The low-abundance elution peaks of each group of samples after removal of high-abundance proteins were collected and combined, and concentrated by ultrafiltration using a 15mL 5kDa ultrafiltration tube, and replaced by buffer with 25mmol / L ammonium bicarbonate solution, keeping the final tot...

Embodiment 3

[0050] Childhood asthma decision tree model for protein combinatorial markers:

[0051] Apolipoprotein C3, apolipoprotein E, and serum amyloid A were analyzed by Biomarker Patterns Software 5.0 software and compared the weight combination of each protein. Figure 7 ). ①When apolipoprotein E≤48.28, and apolipoprotein C3>156.45μg / mL; ②When apolipoprotein E≤48.28μg / mL, apolipoprotein C3≤156.45μg / mL, and serum amyloid A≤ At 0.65 μg / mL, both conditions were judged as healthy children. ①When apolipoprotein E>48.28μg / mL; ②When apolipoprotein E≤48.28μg / mL, apolipoprotein C3≤156.45μg / mL, and serum amyloid A>0.65μg / mL, the two All cases were judged as asthmatic children. The model was used to detect 120 children with asthma and 40 healthy children's serum samples. The sensitivity of the model for childhood asthma was 79.2%, the specificity was 92.5%, the positive predictive value was 96.9%, and the Youden index was 0.717. 10- After fold cross-validation, the model had a sensitivity ...

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Abstract

The invention discloses an application of a protein combined marker in preparation of a children asthma diagnostic reagent. The protein marker comprises protein apolipoprotein C3, apolipoprotein E andserum amyloid protein A. The protein combined type children asthma diagnostic reagent is simple to prepare and simple and easy to sample and detect, is capable of achieving quantitative detection, issmall in sample amount, high in diagnosis sensitivity and high in specificity, and is capable of achieving high-throughput detection.

Description

technical field [0001] The invention belongs to the field of biology, and particularly relates to the application of a protein combination marker related to complement and blood coagulation cascade reaction pathways in the preparation of children's asthma diagnostic reagents. Background technique [0002] Bronchial asthma, short for asthma, is a chronic respiratory disease characterized by persistent airway inflammation, airway hyperresponsiveness, and airway remodeling. The incidence of childhood asthma in my country is increasing year by year, and the overall prevalence of asthma among children aged 0-14 in urban my country has increased by 43.4% compared with 10 years ago. Although 2 / 3 of children with asthma will disappear with age, 1 / 3 of children will develop into adult asthma. [0003] At present, asthma in children is mainly diagnosed and evaluated by clinical symptoms, pulmonary function tests and treatment effects. In terms of clinical symptoms, the clinical mani...

Claims

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Application Information

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IPC IPC(8): G01N33/68
CPCG01N33/6893G01N2800/122G01N2333/775
Inventor 刘姬艳贺琳娟
Owner HANGZHOU NORMAL UNIVERSITY
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