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ELISA kit for detecting human BP180 IgG antibody and application thereof

A BP180 and kit technology, applied in the field of medicine, can solve the problem of not meeting the diagnostic needs of clinical BP and anti-BP180MMP, and achieve the effect of cost increase and cost saving

Pending Publication Date: 2022-03-25
李小光
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Therefore, the current commercial BP180 IgG antibody detection kit that recognizes BP180 NC16a obviously cannot meet the diagnostic needs of clinical BP and anti-BP180 MMP

Method used

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  • ELISA kit for detecting human BP180 IgG antibody and application thereof
  • ELISA kit for detecting human BP180 IgG antibody and application thereof
  • ELISA kit for detecting human BP180 IgG antibody and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] Embodiment 1 is used to detect the preparation of the ELISA kit of BP180 IgG antibody

[0042] 1. Composition:

[0043] ELISA kit includes:

[0044] (1) BP180 microwell strip (48 wells)

[0045] (2) 0 U / ml standard serum 1 (reaction buffer solution containing 0.05% sodium azide w / v, 1.5ml)

[0046] (3) 100U / ml standard serum 2 (serum from patients with positive BP180 IgG autoantibody diluted in reaction buffer containing 0.05% w / v sodium azide, 1.5ml)

[0047] (4) Enzyme-labeled antibody (horseradish peroxidase-labeled rabbit anti-human IgG antibody, 8ml)

[0048] (5) 1x reaction buffer (0.01M PBS containing 0.05% v / v Tween20 and 0.05% w / v NaN3, 50ml)

[0049](6) 10x wash buffer (0.1M PBS containing 0.5% v / v Tween20, 100ml)

[0050] (7) Enzyme matrix solution (ready-to-use TMB chromogenic solution, 8ml)

[0051] (8) Stop solution (0.5N hydrochloric acid solution, 8ml)

[0052] 2. Coating of BP180 microwell strip

[0053] (1) The coated material includes 29 BP180...

Embodiment 2

[0079] The performance index detection of embodiment 2 kit

[0080] (1) Sample source

[0081] Bullous pemphigoid, mucosal pemphigoid (BP180 type), mucosal pemphigoid (anti-laminin 332) and pemphigus vulgaris were collected 50 times each, part of which came from the Department of Dermatology, Kurume University, Japan. Donated by Professor Takashi Hashimoto (collected from patients in Japan and all over the world), the other part comes from the serum of the remaining patients after the serological diagnosis of skin autoimmune blister disease provided by the dermatology department of a tertiary hospital in China or a dermatology hospital . 300 parts of normal human serum were the remaining serum from normal physical examination in the hospital.

[0082] Serum was diluted 1:50.

[0083] (2) Specificity and sensitivity

[0084] 50 parts of bullous pemphigoid, mucosal pemphigoid (BP180 type) patient sera were detected using commercialized ELISA kit (MBL company) and the ELISA k...

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Abstract

The invention discloses an ELISA (enzyme-linked immuno sorbent assay) kit for detecting a human BP180 IgG (immunoglobulin G) antibody and application of the ELISA kit. The kit contains a BP180 microporous band, and the BP180 microporous band is respectively coated with polypeptides as shown in SEQ ID NO. 1-29 or a mixture of the polypeptides as shown in SEQ ID NO. 1-29. Preferably, the kit also contains standard serum 1 and 2, an enzyme-labeled antibody, a diluent, a reaction buffer solution, a cleaning buffer solution, an enzyme matrix solution and a stop solution. According to the invention, a plurality of antigen polypeptides are used for replacing a long-fragment complete protein as a coating protein of ELISA, so that a large amount of cost is saved while antigen epitopes recognized by an antibody are fully guaranteed, and compared with NC16a recombinant protein, the sensitivity is higher. According to the invention, the human BP180 IgG antibody is detected by using an ELISA method, and compared with an existing immunoblotting method, the ELISA method is easier to be applied to clinical diagnosis of bullous pemphigus and BP180 type mucous membrane pemphigus.

Description

technical field [0001] The invention relates to an ELISA kit and its application, in particular to an ELISA kit for detecting human BP180 IgG antibody and its application. The invention belongs to the technical field of medicine. Background technique [0002] Bullous pemphigoid (BP) and BP180-type mucous membrane pemphigoid (anti-BP180-type MMP) both belong to human skin autoimmune blistering diseases, which mainly affect the skin of the elderly. Immune vesicular disease. The similarity between the two is that both need to detect anti-BP180 IgG autoantibodies to make a diagnosis. The difference is mainly in the clinical features. Patients with bullous pemphigoid only have skin lesions, while BP180 mucosal pemphigoid can Presents as damage to the skin and mucous membranes, or only to the mucous membranes. [0003] Existing methods for detecting IgG autoantibodies to human BP180 mainly include immunoblotting and ELISA. Western blotting mainly uses epidermal extracts or rec...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68G01N33/58G01N33/543
CPCG01N33/6854G01N33/6893G01N33/581G01N33/54306G01N2800/24G01N2800/20
Inventor 李小光钱华周志军
Owner 李小光
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