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An antibody chip kit for detecting periodontal disease-related proteins

A related protein and antibody chip technology, applied in biological testing, measuring devices, material inspection products, etc., can solve the problems of cumbersome operation, low sensitivity, and single detection index, and achieve high detection sensitivity, low background, and less specimen consumption Effect

Active Publication Date: 2019-05-21
RAYBIOTECH INC GUANGZHOU
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] The purpose of the present invention is to overcome the deficiencies of the prior art, and provide an antibody chip kit for detecting periodontal disease-related proteins, which can detect multiple periodontal disease-related proteins simultaneously, and overcome the cumbersome operation of the prior art, It has defects such as single detection index and low sensitivity, and has the advantages of cheap, convenient, sensitive, accurate, high-throughput, less specimen consumption, and can be popularized and scaled in ordinary laboratories.

Method used

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  • An antibody chip kit for detecting periodontal disease-related proteins
  • An antibody chip kit for detecting periodontal disease-related proteins
  • An antibody chip kit for detecting periodontal disease-related proteins

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Experimental program
Comparison scheme
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Embodiment 1

[0021] Screening of antibody chip carriers: Conventional antibody chips mostly use nitrocellulose membrane as a carrier. Since the nitrocellulose membrane has a multi-layer structure, it is difficult to wash the chip, resulting in a high background of the chip and large fluctuations in the results. At the same time, the nitrocellulose membrane is fragile, not easy to operate on a large scale, and it is not widely used for large-scale clinical samples. Traditional glass slides are cheap and have low background, which has brought breakthroughs in the widespread use of diagnostic and research fields. We screened the slides on the market that were not treated with the active method, and the spotting effect of the slides that were either aldehyde or aminated was not stable. After a lot of screening, it is found that the temperature and the active reagent components on the surface of the slide are the key to the effect of spotting on the slide.

[0022] The following 5 sets of slid...

Embodiment 2

[0028] Preparation of an antibody chip kit for simultaneous quantitative detection of multiple periodontal disease-related proteins: in order to detect whether there are corresponding receptors in the sample, prepare slides immobilized with specific antibodies corresponding to 20 kinds of periodontal disease-related proteins, The specific antibodies corresponding to the 20 periodontal disease-related proteins are common commercially available products.

[0029] Preparation and storage of the antibody chip: 100-1000 pl of PBS buffer solution containing specific antibodies (containing 0.01-10 g / 100 ml bovine albumin) was spotted on a glass slide with an automatic spotting device. The specific chip array uses biotin-labeled bovine IgG as a positive control. There are four replicate spots in each array for 20 antibodies and two different concentrations of positive controls. In this embodiment, the chip array adopts such as figure 1 The arrangement shown, but in fact, in other em...

Embodiment 3

[0034] The experimental steps of using the kit of the present invention to quantitatively detect various periodontal disease-related proteins are as follows:

[0035] 1. Complete drying of the slide chip: Take the slide chip out of the box, and after equilibrating at room temperature for 20-30 minutes, open the packaging bag, uncover the sealing strip, and then put the chip in a vacuum desiccator or dry at room temperature for 1-20 minutes. 2 hours.

[0036] 2. Gradient dilution of periodontal disease-related proteins according to Table 2

[0037] 2.1. Add 500 μl of sample diluent to the small tube of periodontal disease-related protein standard mixture, and redissolve the standard. Before opening the vial, give it a quick centrifuge and gently pipet up and down to dissolve the powder. Label this vial as Std1.

[0038] 2.2. Mark 6 clean centrifuge tubes as Std2, Std3 to Std7 respectively, and add 200 μl of sample diluent to each small tube.

[0039] 2.3. Take 100 μl of Std1...

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Abstract

Provided is an antibody-array test kit for detecting proteins related to periodontal disease, comprising an antibody array; said antibody array uses as its solid-phase support a glass slide treated with a hydrophilic reagent containing an alkyl glycoside, and a specific antibody mixture of various periodontal disease-related proteins is spotted onto the solid-phase support under conditions of 22-24°C and 30-40% humidity. The test kit is capable of detecting a plurality of proteins related to periodontal disease and commonly used for clinical purposes, and has the advantages that it is sensitive, high-throughput, uses a small amount of specimen, and may be popularized and used on a large scale in conventional laboratories.

Description

technical field [0001] The invention relates to the technical field of biological detection, in particular to an antibody chip kit for detecting periodontal disease-related proteins. Background technique [0002] Periodontal disease refers to diseases that occur in the tooth supporting tissue (periodontal tissue), including gingival disease involving only gingival tissue and periodontitis involving deep periodontal tissue (periodontal ligament, alveolar bone, cementum). category. Periodontal disease is a common oral disease, one of the main causes of adult tooth loss, and a major oral disease that endangers human teeth and general health. The early symptoms of periodontal disease are not easy to pay attention to, resulting in long-term chronic infection of periodontal tissue, and recurrent inflammation, which not only damages the function of the oral masticatory system, but also seriously affects health. [0003] Periodontal disease is a disease that ranges from simple inf...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/68G01N33/573G01N33/543
CPCG01N33/543
Inventor 黄若磐毛应清
Owner RAYBIOTECH INC GUANGZHOU
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