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CN-120703383-B - Kit for detecting allergen-specific IgE antibody

CN120703383BCN 120703383 BCN120703383 BCN 120703383BCN-120703383-B

Abstract

The application relates to the technical field of in-vitro diagnosis, in particular to a kit for detecting allergen-specific IgE antibodies. The kit comprises an anti-interference agent used for being mixed with a sample to be tested, an immune labeled allergen, a peroxidase compound capable of combining with the immune label and catalyzing color development of a substrate, and the substrate capable of being catalyzed by the peroxidase to develop color, wherein the anti-interference agent comprises, by weight, 3-6 parts of polystyrene microspheres, 12-18 parts of L-glucoside and 6-10 parts of casein. The kit provided by the application can effectively reduce false positive and false positive results, thereby improving the accuracy of detection results.

Inventors

  • FENG ZHIQING
  • YANG SUQI

Assignees

  • 北京新华联协和药业有限责任公司

Dates

Publication Date
20260512
Application Date
20250709

Claims (9)

  1. 1. A kit for allergen-specific IgE antibody detection, characterized in that the kit comprises an anti-interference agent for mixing with a sample to be tested, an immunolabeled allergen, a complex of a peroxidase capable of binding to the immunolabel and of catalyzing the development of a substrate, a substrate capable of being catalytically developed by the peroxidase; The anti-interference agent comprises, by weight, 4-5 parts of polystyrene microspheres, 14-16 parts of L-glycoside and 7-9 parts of casein, wherein the amino acid sequence of the casein is a sequence shown as SEQ ID NO 1, and the particle size of the polystyrene microspheres is 50nm-1 mu m.
  2. 2. The kit according to claim 1, wherein the kit further comprises a buffer solution, wherein the buffer solution is 0.01-0.1M Tris-HCl buffer solution with pH of 7.0-7.5.
  3. 3. The kit according to claim 2, wherein the anti-interference agent is mixed with the buffer solution, and the concentration of the anti-interference agent in the mixed solution after mixing is 0.2-0.8mg/mL.
  4. 4. The kit according to claim 3, wherein the anti-interference agent is mixed with the buffer solution, and the concentration of the anti-interference agent in the mixed solution after mixing is 0.4-0.7mg/mL.
  5. 5. The kit of claim 1, wherein the complex of peroxidases capable of binding to the immunological label and capable of catalyzing the development of a substrate comprises streptavidin coupled to the peroxidases.
  6. 6. The kit of claim 1, wherein the immunolabel comprises a biotin label, a FITC label.
  7. 7. The kit of claim 1, wherein the allergen comprises an intake allergen and an inhalation allergen.
  8. 8. The kit of claim 7, wherein the food allergen comprises any one or more of eggs, milk, beans, seafood, dried fruits.
  9. 9. The kit of claim 7, wherein the inhaled allergen comprises any one or more of pollen species, dust mites species, animal dander species, mold species.

Description

Kit for detecting allergen-specific IgE antibody Technical Field The application relates to the technical field of in-vitro diagnosis, in particular to a kit for detecting allergen-specific IgE antibodies. Background In the field of allergen detection, the method for detecting specific IgE in vitro is widely applied, and can effectively assist in diagnosing allergic diseases. However, in the actual detection process, various interference substances exist in the sample, which seriously affect the accuracy of the detection result, so that false positive or false negative results appear and interfere with clinical diagnosis. The types of interfering substances in the sample are numerous. The heterophilic antibody is used as an antibody against an undefined antigen existing in human peripheral circulation blood, has multi-specificity and weak affinity, and can interfere with detection of various indexes such as Human Chorionic Gonadotropin (HCG), follicle Stimulating Hormone (FSH) and the like by combining with an Fc segment of IgG and interacting with a labeled antibody or a capture antibody in a detection system. The rheumatoid factor belongs to autoantibodies, denatured IgG is used as a target antigen, antibodies in a detection system can be bridged, interference is generated on immune detection, and the interference degree and concentration have no definite dose effect relation. Human anti-animal antibodies (such as the common human anti-mouse antibody HAMA), caused by iatrogenic or non-iatrogenic factors, have strong affinity, are specific to antigens, and can interfere with antibodies of corresponding sources. In addition, complement, autoantibodies, hormone binding proteins, etc. in the sample can also interfere with detection. The level of complement rises under the conditions of acute inflammation and the like, which can lead to false positive or false negative results in an enzyme-linked immunosorbent assay (ELISA) detection system, the autoantibody and the target antigen are combined to form a complex to influence the target antigen detection result, and the hormone binding protein and the hormone are combined to change the concentration of an object to be detected in a sample to interfere detection. To solve the above-mentioned interference problem, it is necessary to add a blocking agent to the allergen IgE detection kit. The non-specific blocker plays a role in aiming at a plurality of interference substances widely existing in a sample, prevents non-specific binding by means of saturation of binding sites which are not bound, provision of alternative binding sites, steric hindrance caused by specific binding with the interference substances and the like, and reduces the binding of the interference antibody and the test antibody component, thereby improving the sensitivity and the specificity of detection. Currently, there are many blocking agents available on the market for allergen detection kits, but there are still many problems. Some blocking agents have poor blocking effects on specific interfering substances, and cannot fully cope with various interferences in complex samples. For example, animal IgG is a common passive blocker, and although alternative binding sites can be provided to prevent binding of interfering antibodies to capture antibodies or detection antibodies, only specific types of interference can be blocked, multiple IgG complexes are required and the blocking effect depends on the affinity of the interfering antibodies to the animal IgG. In addition, some blockers can have a negative impact on the detection signal while reducing interference, e.g., excessive use of passive blockers can result in reduced detection signals, affecting detection of low concentrations of allergen IgE. Meanwhile, the applicability of the existing blocking agent in different sample types is different, and the blocking effect can not be ensured to be stably exerted in various samples such as serum, plasma, nasal secretion and the like. Thus, there is a need to further optimize blockers in allergen IgE detection kits. Disclosure of Invention The application provides a kit for allergen-specific IgE antibody detection. The kit provided by the application can effectively reduce false positive and false positive results, thereby improving the accuracy of detection results. In a first aspect, the present application provides a kit for allergen-specific IgE antibody detection, comprising the following technical scheme: A kit for detection of allergen-specific IgE antibodies, the kit comprising an anti-interference agent for mixing with a sample to be tested, an immunolabeled allergen, a complex of a peroxidase capable of binding to the immunolabel and of catalyzing the development of a substrate, a substrate capable of being catalytically developed by the peroxidase; the anti-interference agent comprises, by weight, 3-6 parts of polystyrene microspheres, 12-18 parts of L-glycoside and 6-10 parts of casein