CN-120559258-B - Kit for detecting content of free IgE in human serum and application thereof

Abstract

The application relates to the technical field of in-vitro detection, and particularly discloses a kit for detecting the content of free IgE in human serum and application thereof. The kit comprises a capture reagent, a binding reagent and a magnetic separation reagent, wherein the capture reagent comprises Fc epsilon RI protein and a stabilizer, and the stabilizer comprises, by weight, 0.10-0.50 parts of hyaluronic acid, 0.40-0.80 parts of xanthan gum and 0.15-0.35 parts of sodium lauryl amphoacetate. The kit provided by the application can effectively improve the stability of the kit and the accuracy of the detection result.

Inventors

• FENG ZHIQING
• YANG SUQI

Assignees

• 北京新华联协和药业有限责任公司

Dates

Publication Date

20260508

Application Date

20250703

Claims (6)

1. 1. A kit for detecting the content of free IgE in human serum is characterized by comprising a capture reagent, a binding reagent and a magnetic separation reagent, wherein the capture reagent comprises Fc epsilon RI protein and a stabilizer, and the stabilizer comprises, by weight, 0.10-0.50 part of hyaluronic acid, 0.40-0.80 part of xanthan gum and 0.15-0.35 part of sodium lauryl amphoacetate.
2. 2. The kit according to claim 1, wherein the stabilizer comprises the following components, by weight, 0.20-0.40 parts of hyaluronic acid, 0.50-0.70 parts of xanthan gum, and 0.20-0.30 parts of sodium lauroamphoacetate.
3. 3. The kit according to claim 1, wherein the addition amount of xanthan gum in the stabilizer is 1.5 to 2.5 times the addition amount of hyaluronic acid.
4. 4. The kit according to claim 1, wherein the weight ratio of the fceri protein and the stabilizer is (2-4): 100.
5. 5. The kit according to claim 4, wherein the weight ratio of the Fc epsilon RI protein to the stabilizer is (2.5-3.5): 100.
6. 6. The kit of claim 1, wherein the concentration of fceri protein in the capture reagent is 0.10-0.20ug/mL.

Description

Kit for detecting content of free IgE in human serum and application thereof Technical Field The application relates to the technical field of in-vitro detection, in particular to a kit for detecting the content of free IgE in human serum and application thereof. Background IgE is a key antibody that mediates type I hypersensitivity reactions, and free IgE levels directly reflect the degree of activation of the allergic state of the body. By detecting the concentration, igE-mediated immediate allergy (such as food allergy and anaphylactic shock) and non-IgE-mediated allergy (such as contact dermatitis) can be distinguished, and a basis is provided for accurate treatment. For example, (1) in acute allergic attacks, for suspected anaphylactic shock and laryngeal oedema patients, detection of free IgE can rapidly confirm IgE-mediated mechanism and guide the use of first-aid drugs such as epinephrine, (2) in chronic allergic disease identification, about 30% of chronic urticaria patients are IgE-mediated, detection of free IgE can distinguish the etiology and avoid excessive use of glucocorticoid, and (3) in asthmatic patients, allergic asthma (IgE elevation) needs to be identified from non-allergic asthma (IgE normal) to select anti-IgE treatment (such as omalizumab). At the same time, free IgE levels are positively correlated with allergy severity, e.g. when free IgE >100IU/mL for asthmatic patients, the risk of severe episodes increases significantly, so dynamic monitoring of free IgE levels can guide treatment regimen adjustment. In addition, the continuous rise of free IgE in infancy suggests an increased risk of atopic process, and the possibility of asthma and allergic rhinitis in the future can be predicted. Therefore, the detection of the free IgE content in serum is particularly important. However, at present, there is little to no detection of free IgE in human serum in the clinic. Thus, there is a need to provide a method for detecting the free IgE content in serum. Disclosure of Invention The application provides a kit for detecting the content of free IgE in human serum and application thereof. The kit provided by the application can effectively improve the stability of the kit and the accuracy of the detection result. Fceri proteins are high affinity receptors for IgE and are capable of recognizing and specifically binding to IgE. The detection of the free IgE content is achieved by the specific binding of the Fc epsilon RI protein to IgE. In a first aspect, the application provides a kit for detecting the content of free IgE in human serum, which adopts the following technical scheme: The kit for detecting the content of free IgE in human serum comprises a capture reagent, a binding reagent and a magnetic separation reagent, wherein the capture reagent comprises Fc epsilon RI protein and a stabilizer, and the stabilizer comprises, by weight, 0.10-0.50 part of hyaluronic acid, 0.40-0.80 part of xanthan gum and 0.15-0.35 part of sodium lauroamphoacetate. According to the application, the combination of hyaluronic acid, xanthan gum and sodium lauroamphoacetate is added into the capture reagent as the stabilizer, so that on one hand, compared with the use of the stabilizer in the related technology in the capture reagent of the kit, the detection result of the kit provided by the application has better stability. On the other hand, the kit provided by the application can be used for detecting the sample to be detected, and the accuracy of the detection result can be effectively improved. Hyaluronic acid is a high molecular polysaccharide, and sodium hyaluronate in the form of sodium salt has good water solubility and stability. Xanthan gum is a microbial extracellular polysaccharide produced by fermenting Xanthomonas oleracea (Xanthomonas campestris) and is formed by connecting glucose, mannose and glucuronic acid through glycosidic bonds. The molecular structure of the modified polyurethane contains a large number of negative charge groups. Sodium lauryl amphoacetate is a zwitterionic surfactant. The hydrophobic chain of the sodium lauroamphoacetate can be preferentially adsorbed on the hydrophobic region or the gas-liquid interface of the antibody, so that the hydrophobic aggregation of the antibody and the inactivation caused by interface adsorption (such as loss caused by adsorption of the antibody to the pore walls of the plate in detection) are reduced. Xanthan gum (negative charge) and sodium lauryl amphoacetate (zwitterionic, negatively charged at neutral pH) may enhance system stability through electrostatic repulsion, especially under neutral or weakly alkaline conditions, both negative charges synergistically inhibit antibody aggregate formation. Through experimental analysis, when the kit is used for detecting a sample to be detected, compared with the method that any one or two of hyaluronic acid, xanthan gum and sodium lauroamphoacetate are selected to be added into a capture reagent o