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Chemiluminiscence diagnostic kit for sensitization allergens and preparation method thereof

A diagnostic kit and chemiluminescence technology, applied in chemiluminescence/bioluminescence, analysis through chemical reaction of materials, scientific instruments, etc., can solve problems such as being in the blank, and achieve low diagnostic cost, long validity period, good repeatability

Inactive Publication Date: 2013-04-10
北京新华联协和药业有限责任公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, in the diagnosis of allergens in allergic diseases, there has been a blank

Method used

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  • Chemiluminiscence diagnostic kit for sensitization allergens and preparation method thereof
  • Chemiluminiscence diagnostic kit for sensitization allergens and preparation method thereof
  • Chemiluminiscence diagnostic kit for sensitization allergens and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0053] Embodiment 1 Preparation of Allergen Chemiluminescent Method Diagnostic Kit I

[0054] 1.1 Preparation of chemiluminescent plates coated with allergens

[0055] Preparation of allergenic house dust mite / dust mite, artemisia pollen, ragweed pollen, dog / cat hair, cockroach, humulus pollen, mold mix. Each allergen was treated with 50mM carbonate buffer (Na 2 CO 3 1.59g / L and NaHCO 3 2.93g / L) diluted to 2.5μg / ml, added to the corresponding luminescent plate, 100μl per well, incubated overnight at 4°C, washed the luminescent plate 3 times with phosphate buffer containing Tween 20, and then added phosphate containing BSA Buffer (NaCl 8g / L, KCl 2g / L, NaCl 2 HPO 4 2.87g / L, KH 2 PO 4 0.2g / L, Tween 200.5ml / L, 5%BSA), after standing at room temperature for 2 hours, discard the liquid in the well, dry the luminescent plate thoroughly, vacuum pack it in an aluminum bag, and complete the pre-coated luminescence coated with the allergen Plate preparation.

[0056] Wherein, th...

Embodiment 2

[0074] Example 2 Preparation of Allergen Chemiluminescence Diagnostic Kit II

[0075] 1.1 Preparation of chemiluminescent plates coated with allergens

[0076] Preparation of allergen spring pollen Ⅰ (cedar / fir / poplar / elm / willow), spring pollen Ⅱ (birch / maple / oak / walnut / rape), polyvalent fungi Ⅰ (Penicillium chrysogenum / black Aspergillus / Trichoderma konii / Mucor racemosa / Rhizopus stoloni), polyvalent fungi Ⅱ (Peophysis / Curvularia / Cladosporium macrosporum / Helminthes sp. Herbivorous smut / Smut sativa / Fusarium graminearum / Smut smut / Cephalosporium), tree pollen combination (willow / poplar / juniper / sycamore / white wax / elm). Dilute each allergen with 50mM carbonate buffer (Na 2 CO 3 1.59g / L and NaHCO 3 2.93g / L) to 5μg / ml, add to the corresponding luminescent plate, 100μl per well, incubate overnight at 4°C, wash the luminescent plate 3 times with phosphate buffer containing Tween 20, and then add phosphate buffer containing BSA solution (NaCl 8g / L, KCl 2g / L, NaCl 2 HPO ...

Embodiment 3

[0086] Example 3 Preparation of Allergen Chemiluminescence Diagnostic Kit III

[0087] 1.1 Preparation of chemiluminescent plates coated with allergens

[0088] Preparation of allergens Summer and Autumn Pollen Ⅰ (Sunflower / Cocklebur / Ashweed / Hemp), Summer and Autumn Pollen Ⅱ (Corn Pollen / Sorghum Pollen / Sedge Pollen / Castor Bean Pollen), Artemisia Pollen, Humulus Pollen, Ragweed Pollen, Polyvalent fungi Ⅰ (Penicillium chrysogenum / Aspergillus niger / Trichoderma konii / Mucor racemosa / Rhizostrichum), polyvalent fungi Ⅱ (Piophyllum sporogenes / Curvularia sp. fungus / baker's yeast), polyvalent fungi Ⅲ (Chemophagocystis sp. Each allergen was treated with 50mM carbonate buffer (Na 2 CO 3 1.59g / L and NaHCO 3 2.93g / L) diluted to 5μg / ml, added to the corresponding luminescent plate, 100μl per well, incubated overnight at 4°C, washed the luminescent plate 3 times with phosphate buffer containing Tween 20, and then added phosphate buffer containing BSA solution (NaCl 8g / L, KCl 2g / L, NaCl ...

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Abstract

The invention provides a chemiluminiscence diagnostic kit for sensitization allergens and a preparation method thereof. The diagnostic kit comprises a microporous plate coated with the sensitization allergens, a horseradish peroxidase-labeled anti-human IgE (immunoglobulin e) antibody, chemiluminiscence substrate solutions A and B, and a washing solution, wherein the sensitization allergens comprise one or more of an insect allergen, a pollen allergen, a fungus allergen and a food allergen. According to the diagnostic kit, the difference of the sensitivities of different proteins in one diagnostic kit is overcome, and the detection sensitivity of each allergen is enhanced to the maximum extent by repeatedly testing and adjusting the coating amount of the allergens, the concentration of the enzyme-labeled antibody and the formulae and the concentrations of the chemiluminiscence substrate solutions. The chemiluminiscence diagnostic kit is higher in detection sensitivity, safe, reliable, simple and convenient to operate and low in cost in comparison with the ELISA (enzyme linked immunosorbent assay), western-blot and RAST (radioallergo-sorbent test) diagnostic kits for the allergens.

Description

technical field [0001] The invention relates to the field of detection of allergens, in particular to a chemiluminescence diagnostic kit for allergens and a preparation method thereof. Background technique [0002] Allergic diseases (also known as allergic diseases) include atopic dermatitis, food allergy, allergic rhinitis and allergic asthma, etc., and their incidence is increasing day by day, and the condition is becoming more and more complicated. WHO has listed allergic diseases as key research and prevention diseases in the 21st century. In recent years, with the development of immunohistochemistry, molecular biology techniques and clinical new technologies, such as fiberoptic bronchoscopy, a consensus has been reached on the disease, that is, it belongs to allergic inflammation, and there are a large number of inflammatory cells (including eosinophils) in the inflammatory area. cells, lymphocytes, mast cells, basophils, etc.). The pathogenesis of the disease mainly ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/543G01N21/76
Inventor 白彩明姜敏裴潇竹汤承祁牛占坡马博晨
Owner 北京新华联协和药业有限责任公司
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