BP180 IgE antibody concentration detection kit and detection method and application thereof

A technology for detecting kits and antibody concentrations, applied in the field of BP180IgE antibody concentration detection kits, which can solve the problems of disease severity assessment and no NBP-specific detection methods.

Pending Publication Date: 2020-10-23
李丽沙 +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

But so far, there are very few studies on NBP, and there is no repo

Method used

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  • BP180 IgE antibody concentration detection kit and detection method and application thereof
  • BP180 IgE antibody concentration detection kit and detection method and application thereof
  • BP180 IgE antibody concentration detection kit and detection method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

preparation example Construction

[0045] Secondly, this protocol provides a reagent preparation method for detecting BP180 IgE antibody detection ELISA kit in serum, including:

[0046] 1) Prokaryotic expression of BP180 NC16A antigen

[0047] (1) Preparation of Competent Cells

[0048] ① After LB solid culture and autoclaving, cool to about 60°C, add ampicillin to make the final concentration reach 100 μg / mL, shake well and plate.

[0049] ②Pick a single colony from the reactivated E.coli DH5α plate, inoculate it in 3-5mL LB liquid medium, culture at 37°C with shaking at 220rmp until logarithmic growth (A600 0.6-0.8 measured by spectrophotometer, about 12h).

[0050] ③Transfer the bacterial suspension into 100mL LB liquid medium at a ratio of 1:100 to 1:50, and incubate with shaking at 37°C and 220rmp for about 3 hours until A600=0.5.

[0051] ④Transfer the culture solution into a centrifuge tube (8mL / tube), place on ice for 10-20 minutes, and then centrifuge at 3000g for 10min at 4°C.

[0052] ⑤ Discard t...

Embodiment

[0134] Example: Study on the correlation between BP180 IgE antibody level and NBP condition

[0135] 1. Research object

[0136] The 35 cases of typical BP patients and 30 cases of NBP patients included in this study were first-time diagnosed patients in the Department of Dermatology of the First Affiliated Hospital of Dalian Medical University. They were all in the active stage of the disease and before receiving any systemic treatment. All patients were diagnosed by experienced dermatologists based on clinical manifestations, histopathological examinations, and direct and indirect immunofluorescence examinations and met the BP diagnostic criteria established by the British Skin Association in 2012. For each patient, after being informed of the purpose of the study, he agreed to donate his blood for this study and signed an informed consent form. Patients with pregnancy, other autoimmune diseases, other skin diseases, asthma, parasitic infections, hypereosinophilic syndrome,...

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Abstract

The invention provides a BP180 IgE antibody concentration detection kit and a detection method and an application thereof and belongs to the technical field of IgE detection methods. The kit comprisesa microporous strip, standard serum I, standard serum II, an enzyme labeled antibody solution, a buffer solution for reaction, a cleaning buffer solution, an enzyme matrix solution, a stop solution,a sealing plate and a pasting film. According to the BP180 IgE antibody concentration detection kit and the detection method and the application thereof provided by the invention, rapid detection of the BP180 IgE antibody in serum of an NBP patient is greatly facilitated, meanwhile, the concentration of the BP180 IgE antibody in the serum of a patient can be rapidly and effectively detected, so the concentration of the BP180 IgE antibody is used as one of secondary indexes of NBP diagnosis, furthermore, the BP180 IgE antibody in serum of a patient is detected by the BP180 IgE antibody concentration detection kit, and the BP180 IgE antibody concentration detection kit can further be used as one of indexes for evaluating the severity degree of the NBP disease.

Description

technical field [0001] The present invention relates to the technical field of IgE detection methods, in particular to a BP180 IgE antibody concentration detection kit and its detection method and application. Background technique [0002] Bullous Pemphigoid (BP) is the most common autoantibody-mediated autoimmune bullous skin disease, usually occurs in the elderly, the course of the disease is protracted, severe patients can involve a large area of ​​the body, and die The rate is about 30%-50%. The typical clinical presentation of BP is the appearance of tense vesicles or bullae on the basis of normal skin or erythema, with severe pruritus. However, in recent years, it has been found that more than 20% of BP patients do not have typical bullae, but show itchy non-bullous lesions, such as erythema, urticaria-like plaques, papules, nodules, epidermal exfoliation, eczema, and erythema. illness etc. Therefore, the current type of BP with non-bullous skin lesions as the main ...

Claims

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Application Information

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IPC IPC(8): G01N33/68G01N33/535
CPCG01N33/6854G01N33/535G01N2800/24
Inventor 宋智琦吴畏钟亚男李天博
Owner 李丽沙
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