Dermatophagoides oterronyssinus-specific IgE semi-quantitative detection kit and use thereof

A semi-quantitative detection and kit technology, applied in the field of immunoassay, can solve the problems of complicated steps and long time-consuming, and achieve the effects of simple operation, rapid detection and good correlation

Inactive Publication Date: 2017-10-13
HARBIN MEDICAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] The purpose of the present invention is to overcome the problems of time-consuming, complicated steps, special instruments and other problems in the current detection method of house dust mite sIgE, and provide a semi-quantitative detection of house dust mite sIgE based on fluorescent quantum dot immunochromatography test paper and image analysis technology methods and kits

Method used

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  • Dermatophagoides oterronyssinus-specific IgE semi-quantitative detection kit and use thereof
  • Dermatophagoides oterronyssinus-specific IgE semi-quantitative detection kit and use thereof
  • Dermatophagoides oterronyssinus-specific IgE semi-quantitative detection kit and use thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0061] Example 1: Preparation of calibration quantum dot fluorescent antibody markers

[0062] A. Take 100 μl of quantum dot nanospheres, disperse them in 200 μl of phosphate buffer (10 mM, pH 6.0), centrifuge at 10,000 rpm for 10 min, carefully discard the supernatant, and then add 200 μL of phosphate buffer (10 mM, pH 6.0), Uniformly disperse to obtain quantum dot nanosphere solution;

[0063] B. Weigh 1.9 mg of EDC, dissolve in 1 mL of phosphate buffer, and mix well to obtain a 10 mM EDC solution;

[0064] C. Take 20 μl of EDC solution, add it to 200 μl of quantum dot nanosphere solution, mix well, and incubate at room temperature for 0.5 h;

[0065] D. The activated quantum dot nanospheres were centrifuged at 10,000rpm for 10min, the supernatant was discarded, and then 200μl of boric acid buffer (10mM, pH8.0) was added, and dispersed evenly by ultrasonication;

[0066] E. Take 50 μl of mouse anti-rabbit IgG, add it to the activated quantum dot nanosphere solution, mix we...

Embodiment 2

[0069] Embodiment 2: Preparation of quantum dot-anti-IgE fluorescent antibody marker

[0070] A. Take 100 μl of quantum dot nanospheres, disperse them in 200 μl of phosphate buffer (10 mM, pH6.0), centrifuge at 10,000 rpm for 10 minutes, carefully discard the supernatant, then add 200 μL of phosphate buffer, and disperse evenly to obtain quantum dot nanospheres. ball solution;

[0071] B. Weigh 1.9 mg of EDC, dissolve in 1 mL of phosphate buffer, and mix well to obtain a 10 mM EDC solution;

[0072] C. Take 20 μl of EDC solution, add it to 200 μl of quantum dot nanosphere solution, mix well, and incubate at room temperature for 0.5 h;

[0073] D. The activated quantum dot nanospheres were centrifuged at 10,000rpm for 10min, the supernatant was discarded, and then 200μl of boric acid buffer (10mM, pH8.0) was added, and dispersed evenly by ultrasonication;

[0074] E. Take 50 μl of goat anti-human IgE, add it to the activated quantum dot nanosphere solution, mix well, and then...

Embodiment 3

[0078] Embodiment 3: the assembly of chromatography test strip

[0079] A. Take 100 μl of dust mite allergen protein and dilute it with 100 μl of PBS buffer;

[0080] B. Spray 0.4-1.2mg / mL dust mite allergen protein and rabbit IgG (1mg / mL) on the NC film respectively by using Bio-Dot XYZ 3000 automatic film spotting instrument, as a test strip detection line (T line) With the quality control line (C line), dry overnight at 37°C.

[0081] C. The sample pad, the NC film sprayed with the detection line (T line) and the quality control line (C line), and the water-absorbing pad are sequentially connected and assembled above the plastic substrate used to provide the assembly platform. The assembly schematic diagram is as follows figure 1 shown.

[0082] D. Treat the sample pad with three sample pad treatment solutions A, B, and C respectively. The treatment method is to soak the sample pad in the sample pad treatment solution for 40 minutes, take it out and dry it in a 37°C dryin...

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Abstract

The invention discloses a semi-quantitative detection kit for house dust mite-specific IgE and its application. The test kit includes a detection fluorescent liquid and an immunochromatographic test strip for semi-quantitative detection of house dust mite-specific IgE; wherein, the detection fluorescent liquid is obtained by adding carboxyl-containing quantum dot nanospheres on the surface It is covalently coupled with goat anti-human sIgE and mouse anti-rabbit IgG, and prepared after mixing; the immunochromatography test strip includes sequentially connected sample pads, nitrocellulose membranes, water-absorbing pads, and an assembly platform for providing On the lower plastic backing, the nitrocellulose membrane has a test line coated with dust mite allergen protein and a quality control line coated with rabbit IgG, and the sample pad provides a place for the sample to be tested to be added. The semi-quantitative detection of house dust mite-specific IgE can be realized by using the kit established by the invention, which has a good correlation with clinical symptoms, high detection sensitivity, easy to use, and is suitable for large-scale population allergen screening and family detection Wait for the environment.

Description

technical field [0001] The invention relates to a kit for detecting house dust mite-specific IgE and a detection method thereof, in particular to a house dust mite-specific IgE semi-quantitative detection kit and its method based on fluorescent quantum dot immunochromatography test paper and image analysis technology Detection method. The invention belongs to the technical field of immune analysis. Background technique [0002] House dust mite is one of the most important inhalant allergens, and it is ubiquitous in indoors. House dust mite and farinae farinae are the most important two types of mites. Among them, House Dust mite (HDM) is Oblong, with a body length of about 240-280 μm, also known as European dust mite, is the hottest allergenic mite species in Eurasia. Surveys in Shanghai, Beijing, Guangzhou, Harbin and other places have shown that household dust mites are also the dominant mite species in our country, and they exist on beds, quilts, pillows, carpets, sofas...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68G01N33/558
CPCG01N33/6854G01N33/558G01N33/6893G01N2800/122
Inventor 赵岩孟庆丰安立新
Owner HARBIN MEDICAL UNIVERSITY
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