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Detection kit for basophil activation and method of using the same

A technology of basophils and detection kits, which is applied in the direction of measuring devices, individual particle analysis, particle and sedimentation analysis, etc., can solve the problems affecting the activation results, etc., to avoid activation pathways, comprehensive detection, avoid missed detection and The effect of multiple detection

Active Publication Date: 2017-08-22
HOSPITAL AFFILIATED TO GUANDONG MEDICAL COLLEGE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] In addition, the activation of basophils and its detection process are affected by many factors, such as the serum collection process (choice of anticoagulant), whole blood or isolated white blood cells, whether to add IL-3 and its concentration, allergens (concentration and time), erythrocyte lysate (type) (Pan Qingjun et al., J Immunoassay Immunochem.35:368-377,2014; Pichler WJ(ed):Drug Hypersensitivity.Basel,Karger,2007,391-402), selection of identification parameters , the setting of the activation threshold (the degree of change of the activation index), etc. These factors may affect the judgment of the activation result, and then the diagnosis of clinical diseases

Method used

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  • Detection kit for basophil activation and method of using the same
  • Detection kit for basophil activation and method of using the same
  • Detection kit for basophil activation and method of using the same

Examples

Experimental program
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Embodiment 1

[0039] Example 1: Detecting the activation of basophils in patients with pollen allergy

[0040] An embodiment of the detection kit for basophil activation of the present invention, the detection kit for basophil activation described in this embodiment includes a washing solution, a stimulus control substance, and a basophil with a fluorescent marker. Granulocyte recognition antibody, erythrocyte lysate, antibody staining solution. Among them, the washing solution is 1×PBS, and each 1L washing solution contains: 42.5g NaCl, 1.0gKCl, 0.3g CaCl 2 , 0.3g MgCl 2 .6H 2 O, 1.42gNa 2 HPO 4 .2H 2 O, 0.2g KH 2 PO 4 ;

[0041] Stimulant control substances include positive control substance Ⅰ, positive control substance Ⅱ, allergen and negative control substance; positive control substance Ⅰ is 0.05ng / mL anti-human FcεR Iα antibody, positive control substance Ⅱ is 1nmol / L fMLP, The allergen is the pollen allergen extract (composed of pollen extracts of Chongyang wood, coconut, a...

Embodiment 2

[0057] Example 2: Detection of basophil activation in patients allergic to β-lactam antibiotics

[0058] An embodiment of the detection kit for basophil activation of the present invention, the detection kit for basophil activation described in this embodiment includes a washing solution, a stimulus control substance, and a basophil with a fluorescent marker. Granulocyte recognition antibody, erythrocyte lysate, antibody staining solution. Among them, the washing solution is 1×PBS, each 1L washing solution contains: 8.5g NaCl, 0.2gKCl, 0.5g CaCl 2 , 0.5g MgCl 2 .6H 2 O, 7.1gNa 2 HPO 4 .2H 2 O, 1.0g KH 2 PO 4 ;

[0059] Stimulant control substances include positive control substance Ⅰ, positive control substance Ⅱ, allergen and negative control substance; positive control substance Ⅰ is 10ng / mL anti-human FcεR Iα antibody, positive control substance Ⅱ is 50nmol / L fMLP, allergic It was originally a β-lactam antibiotic allergen (composed of penicillins, cephalosporins, m...

Embodiment 3

[0076] Example 3: Detecting the activation of basophils in patients with autoimmune diseases (autoantigen allergy)

[0077] An embodiment of the detection kit for basophil activation of the present invention, the detection kit for basophil activation described in this embodiment includes a washing solution, a stimulus control substance, and a basophil with a fluorescent marker. Granulocyte recognition antibody, erythrocyte lysate, antibody staining solution. Among them, the washing solution is 1×PBS, and each 1L washing solution contains: 25.5g NaCl, 0.6gKCl, 0.1g CaCl 2 , 0.1g MgCl 2 .6H 2 O, 4.2gNa 2 HPO 4 .2H 2 O, 0.6g KH 2 PO 4 ;

[0078] Stimulant control substances include positive control substance Ⅰ, positive control substance Ⅱ, allergen and negative control substance; positive control substance Ⅰ is 5ng / mL anti-human FcεR Iα antibody, positive control substance Ⅱ is 100nmol / L fMLP, allergic The original is self-antigen (use Thermo Scientific NEPER nuclear pr...

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Abstract

The invention provides a kit for detecting basophil activation and a using method of the kit. The kit comprises a scrubbing solution, an irritant reference substance, fluorescently-labeled basophil recognition antibodies, red blood cell lysis buffer and an antibody staining solution, wherein the fluorescently-labeled basophil recognition antibodies comprise the following basophil recognition antibodies: Anti human CD 11b, Anti human CD 123, Anti human CD 203c and Anti human CD 63; different basophil recognition antibodies have different fluorescent markers. After the kit is used for detecting the basophil activation situation, the known influence factors of the basophil activation and detection can be avoided, the basophil activation situation can be comprehensively and accurately identified and detected, and a better service is provided for the detection of the basophil activation in the aspect of clinical application and scientific research.

Description

technical field [0001] The invention relates to a detection kit for basophil activation and a use method thereof, in particular to a flow cytometry detection kit for basophil activation and a use method thereof. Background technique [0002] At present, the incidence of allergic diseases is increasing year by year in the world. Especially in developing countries, allergic diseases caused by environmental pollution and food hygiene have gradually attracted everyone's attention. Such diseases occupy medical resources The ratio increased month-on-month and significantly affected the quality of life of patients. [0003] Basophils have been known for more than 130 years and are the main effector cells of allergic diseases (Pan Qingjun et al., Chinese Journal of Immunology, 2009, 23(1):25-28). Studies have found that activation is a key step for basophils to exert their biological functions. The study of its activation mechanism has also developed from the classic IgE-mediated a...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N15/14
Inventor 潘庆军刘华锋刘伟敬彭浩晟叶玲冯永民邓真真李璐唐培兰巧芬吴静陈燕文
Owner HOSPITAL AFFILIATED TO GUANDONG MEDICAL COLLEGE