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Joint detection method for rheumatoid factors (IgM type) and other antigen-specific IgM antibodies

A rheumatoid factor, specific technology, applied in the direction of measuring devices, biological tests, material inspection products, etc., can solve the problems of inaccurate results and the inability to realize the detection of rheumatoid factor, etc.

Active Publication Date: 2016-09-28
THE NAT CENT FOR NANOSCI & TECH NCNST OF CHINA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

There have been many literature reports on the use of colloidal gold test strips to detect IgM antibodies, some of which did not deal with the interference of IgG and rheumatoid factors, which may easily cause inaccurate results; some simply increased the ability to absorb human IgG and rheumatoid factors An adsorption line or an adsorption treatment step, without adding the detection line of rheumatoid factor, the detection of rheumatoid factor cannot be realized

Method used

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  • Joint detection method for rheumatoid factors (IgM type) and other antigen-specific IgM antibodies
  • Joint detection method for rheumatoid factors (IgM type) and other antigen-specific IgM antibodies
  • Joint detection method for rheumatoid factors (IgM type) and other antigen-specific IgM antibodies

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0082] This example is used to illustrate the verification method and results of the highly specific anti-human IgM antibody selected in the present invention.

[0083] Method 1 includes: Purified human IgG with a concentration of 1ug / ml, 100ul / well coated in a polystyrene microwell plate at 4 overnight; use 3% bovine serum albumin (BSA), 200ul / well, 37 degrees for 2 hours Blocking; then add anti-human IgM-HRP, 100ul / well, 37 degrees for 30 minutes, after washing, add TMB chromogenic solution, and detect OD with a microplate reader after 10 minutes in the dark 450nm-630nm ; And make positive control wells and blank control wells coated with human IgM; they are all multi-well parallel, and calculate the mean value. The test results are shown in Table 1.

[0084] Method 2 includes: the test strip includes a sample pad, a conjugate pad, a nitrocellulose membrane and a water-absorbent pad that are attached sequentially on a polyvinyl chloride rubber sheet; the sample pad is a gla...

Embodiment 2

[0091] This example is used to illustrate the joint detection test strip and kit of rheumatoid factor (IgM type) and other antigen-specific IgM antibodies and the preparation method thereof of the present invention.

[0092] Such as figure 2 As shown, the test strip of the present embodiment comprises a sample pad, a conjugate pad, a nitrocellulose membrane and a water-absorbent pad attached successively on a polyvinyl chloride rubber sheet; the sample pad is a glass fiber membrane; the conjugate The pad is a glass fiber membrane with an anti-human IgM antibody-colloidal gold conjugate; the nitrocellulose membrane is respectively coated with a first detection line, an adsorption line, a detection line and a quality control line; the first detection line The line is denatured rabbit / human IgG, the adsorption line is an antibody against human IgG, the quality control line is human IgM, and the detection line is the purified Mycoplasma pneumoniae-specific natural multi-epitope a...

Embodiment 3

[0102]This embodiment is used to illustrate the application of the test strip or kit prepared in Example 2 in the detection of RF (IgM type) and Mycoplasma pneumoniae samples.

[0103] The steps of using the test strip are as follows:

[0104] (1) Collect serum samples;

[0105] (2) Put the test strip flat on the detection table, add 100 μl of the serum sample in step (1) dropwise to the sample pad, observe the color development results of the first detection line, detection line and quality control line after 10 to 15 minutes, The color development result after 30 minutes is considered invalid;

[0106] (3) Judgment of results: Judgment results according to Table 3, where "+" means that the red band is displayed, "-" means that the red band is not displayed, and " / " means that the red band is displayed or not:

[0107] table 3

[0108]

[0109] The test strips prepared in Example 2 were used to detect clinically confirmed positive and negative sera, and the results are ...

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Abstract

The invention provides a joint detection test strip for rheumatoid factors (IgM type) and other antigen-specific IgM antibodies. The test strip comprises a sample pad, a conjugate pad, a nitrocellulose membrane and an absorbent pad which are sequentially attached to a polyvinyl chloride rubber slab, wherein a first detection line, an adsorption line, one or more detection lines and a quality control line are sequentially and respectively coated on the nitrocellulose membrane; the first detection line is a denatured rat / human IgG; the adsorption line is an anti-human IgG antibody; the quality control line is human IgM; and the one or more detection lines are respectively selected from one or more specific antigens: a mycoplasma pneumoniae specific antigen, an influenza B specific antigen, a toxoplasma pathogen specific antigen, an autoimmune antigen and allergens. The invention also provides a kit comprising the test strip, a method for preparing the test strip, a detection method by using the test strip or kit, and application of the test strip or kit.

Description

technical field [0001] The invention relates to a test strip. Specifically, the present invention relates to a joint detection test strip for rheumatoid factor (IgM type) and other antigen-specific IgM antibodies, and a test kit including the test strip, a method for preparing the test strip, and using the test strip The method for detecting the test strip or kit, and the application of the test strip or kit. Background technique [0002] Rheumatoid factor (RF) is an autoantibody that targets denatured immunoglobulin G (IgG) and has no species specificity. Rheumatoid arthritis (RA) patients and about 50% of healthy people have RF-producing B cell clones, which can be synthesized and produced in large quantities under the direct action of certain pathological factors such as denatured IgG or Epstein-Barr virus. RF has poor binding ability to natural IgG, but it is easy to react with IgG or polymerized IgG in immune complexes. There are five types of RF: IgG, IgA, IgM, IgD,...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/558G01N33/68
CPCG01N33/558G01N33/68G01N33/6854G01N2800/102
Inventor 蒋兴宇张晓青
Owner THE NAT CENT FOR NANOSCI & TECH NCNST OF CHINA
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