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Quantitative house dust mite allergen specificity IgE antibody detection kit and preparation method thereof

A quantitative detection and allergen technology, applied in measurement devices, analysis, instruments and other directions by causing chemical reaction of materials, can solve the problems of poor high specificity, poor detection sensitivity and accuracy, and achieve high sensitivity and operation. Convenience and accuracy

Inactive Publication Date: 2014-11-26
JIANGSU FLON BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] At present, the ELISA method commonly used in clinical practice, due to its limitations, the detection sensitivity and accuracy are far inferior to the chemiluminescence method, and we have overcome the disadvantage of high background specificity caused by the heterogeneous reaction of ELISA

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0057] A kit for quantitative detection of house dust mite allergen-specific IgE antibodies, said kit containing the following components: magnetic separation reagents, allergen reagents, enzyme reactants, calibration products, quality control products, concentrated cleaning solution, substrate A solution and substrate B solution; wherein, the allergen reagent contains biotin-labeled specific allergen, the magnetic separation reagent contains magnetic particles coated with anti-human IgE antibody, and the substrate A solution contains luminol reagent , the substrate B solution is a peroxide-containing reagent, and the enzyme reactant is a horseradish peroxidase-labeled streptavidin reagent. The concentrated cleaning solution is a concentrated cleaning reagent of phosphate buffer saline containing Tween.

Embodiment 2

[0059] A kind of house dust mite allergen-specific IgE antibody quantitative detection kit described in the above-mentioned embodiment 1, its preparation method is as follows:

[0060] Step 1, Preparation of Magnetic Separation Reagent

[0061] 1. Preparation of magnetic particle buffer

[0062] (1) Weigh TRIS (trishydroxymethylaminomethane) 4.58g / L, NaCl 6.81g / L, TRITON X-100 (polyethylene glycol monooctyl phenyl ether) 0.5ml / L in a 1L container ; (2), measure 0.5-1ml of preservative Proclin-300 in a 1L container; measure 800ml of purified water in the above container, and stir thoroughly; (3), adjust the pH value to 7.95-8.05; (4), Weigh 3-5g of bovine serum albumin BSA into the above-mentioned 1L container; (5), finally dilute to 1000ml, filter through a 0.22μm microporous filter; mark it and store it in a cold room at 2-8°C.

[0063] 2. Preparation of Magnetic Separation Reagents

[0064] (1) Dissolve 1.0 mg disuccinimidyl suberate in 50 μl DMSO, and dissolve 2 mg anti-...

Embodiment 3

[0087] The method of using the house dust mite allergen-specific IgE antibody quantitative detection kit prepared in Example 2 is as follows:

[0088] (1) Add 50 μl of magnetic separation reagent to the bottom of the chemiluminescence reaction tube;

[0089] (2) Add 50 μl of standard substance, quality control substance, and serum to be tested to the bottom of the chemiluminescence reaction tube;

[0090] (3) Gently shake the reaction tube with a multi-functional homogeneous mixer for 30 seconds, then place it in a 37°C incubator for 15 minutes;

[0091] (4) Put the reaction tubes on the magnetic separator, make sure that the bottom of each tube is in contact with the surface of the separator, and settle for 2-3 minutes; slowly invert the separator and pour out the supernatant, and forcefully drain the contents of the reaction tubes on absorbent paper. pat dry to remove droplets sticking to the reaction tube;

[0092] (5) After diluting the concentrated cleaning solution 20 ...

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Abstract

The invention relates to a quantitative detection kit for detecting one or more allergen specificity IgE antibodies as well as a preparation method and a detection method thereof. The preparation method comprises six steps of preparation of a magnetic separation reagent, preparation of an allergen reagent, preparation of an enzyme reactant, preparation of a calibration material and a quality control material, preparation of a wash concentrate and preparation of substrate A liquid and substrate B liquid. The detection method of the quantitative allergen specificity IgE antibody detection kit is implemented by capturing allergens via allergen specificity IgE antibodies; meanwhile, a chemiluminiscence immunological technique and a magnetic separation technique are combined, so that the detection sensitivity and the specificity of the allergen specificity antibodies are greatly improved, and the detection result is relatively real and reliable, the reaction time and the reaction steps are optimized at the same time and the operation is relatively easy.

Description

technical field [0001] The invention relates to a detection kit for house dust mite allergen-specific IgE antibody (quantitative) and a preparation method thereof, which are used for quantitatively measuring the content of IgE antibody specific to house dust mite allergen in human serum in vitro. Background technique [0002] House dust mite is a common inhalant allergen (therefore, the detection of house dust mite-specific IgE antibody is usually called the detection of house dust mite allergen), and the allergic symptoms it can cause are mostly respiratory related diseases. Its detection results are closely related to the improvement of our daily life. [0003] Dust mites are important allergens that induce allergic diseases such as allergic asthma, allergic rhinitis, and eczema-allergic conjunctivitis. Studies in recent years have confirmed that dust mites are closely related to allergic diseases. According to the surveys in many countries such as the United States, New ...

Claims

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Application Information

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IPC IPC(8): G01N33/558G01N21/76
CPCG01N33/54333G01N33/6803G01N2333/4716
Inventor 叶兴旺奚伟红
Owner JIANGSU FLON BIOTECH
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