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Kit for detecting vasculitis related autoantibody repertoire

An autoantibody and antibody detection technology, applied in the biological field, can solve the problems of easy misjudgment and missed judgment, cannot be used as a basis for diagnosis, low specificity, etc., and achieves the effect of uniform width and simple and reliable result judgment.

Active Publication Date: 2013-02-27
SICHUAN XINCHENG BIOLOGICAL CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, it has the following disadvantages: it cannot be used as a basis for diagnosis; the judgment of the result requires rich experience; the significance of titer is greater than that of karyotype, but the judgment of titer is highly subjective; the sensitivity is low and the specificity is not high
[0007] 1. WB is used for a long time and the operation is complicated
[0008] 2. WB uses natural mixed antigens. During the experiment, the problem of not finding the target band and band offset often occurs, which causes many difficulties in the interpretation of the results, and it is easy to misjudge and miss.
[0009] 3. The ELISA method can only detect one item at a time, and the efficiency is not high
[0010] 4. The IFA method can only be used for screening and does not have the significance of auxiliary diagnosis
[0011] 5. There is currently no improved immunoblotting method that can simultaneously detect the three autoantibodies associated with vasculitis
[0012] 6. In the current kits, there is generally no control line or a control line can only be used as a control for one test result, and there is no quality control strip that can be used as a control for at least two test results at the same time

Method used

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  • Kit for detecting vasculitis related autoantibody repertoire
  • Kit for detecting vasculitis related autoantibody repertoire
  • Kit for detecting vasculitis related autoantibody repertoire

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] see Figure 1 to Figure 2 as shown, figure 2 The CO quality control line in the sample is the critical quality control zone; the kit for detecting vasculitis-related autoantibody spectrum in this embodiment includes membrane strips, enzyme labeling solution, substrate and concentrated washing incubation solution, wherein the enzyme labeling solution, substrate The selection and scope of the concentrated washing and incubation solution all belong to the prior art. For the enzyme label solution, the substrate and the concentrated washing incubation solution, the technical scheme of the present invention can also be realized by selecting the products of the prior art.

[0036] The membrane strip is composed of slides and antigen strips, critical quality control strips, and functional quality control lines that are sequentially fixed on the slides. The antigen strips are drawn from at least two independent lines of PR3, MPO and GMB to nitrocellulose Formed on a membrane o...

Embodiment 2

[0044] This example is an experimental determination of the antigen bands formed by three independent streaks of PR3, MPO and GMB on nitrocellulose or nylon membranes. On the basis of Example 1, in order to make the result judgment more simple and reliable, the width of each antigen strip is uniformly set, and the interval between the strips is equal. The width of the antigen strip is 0.5mm-3mm, and the interval between adjacent antigen strips 2mm-20mm, the background of the membrane strip is clean, and an improved single-reagent substrate is used, which does not need to be terminated after adding the substrate. Reagents; all imprinted antigens are highly purified, and the latest international coating technology is used. The specific coating method is described later.

[0045] Determination of critical quality control values:

[0046] 1) Determine the critical quality control value of each antibody

[0047] Randomly select 123 fresh serum and plasma samples clinically diagno...

Embodiment 3

[0056] This embodiment is the critical quality control value of the membrane strip determined according to embodiment 2 (in figure 2 and in all examples of the present invention, CO p is the critical quality control value of a single antibody, CO is the critical quality control value of the entire membrane strip), to adjust the concentration of human IgG coated in the critical quality control zone, and finally determine the coating concentration and coating process of the critical quality control zone. The specific operation is: dissolve a certain concentration of human IgG in Tris or Hepes buffer, then use a fully automatic spotting instrument to streak on the nitrocellulose membrane, and prepare finished membrane strips through sealing, drying, cutting and other processes. And only the critical quality control band is coated on the film strip. Randomly select 30 membrane strips to test the random negative samples of step 1), and scan the gray scale, and calculate the mean ...

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Abstract

The invention relates to a kit for detecting vasculitis related autoantibody repertoire, which comprises a film strip, ELISA (Enzyme Linked Immunosorbent Assay) solution, a substrate and concentrated washing and incubation solution. The film strip consists of a piece of slide glass, and antigen strips, cut-off control strips and a function control line which are sequentially and fixedly arranged on the slide glass; and each antigen strip is formed by separately marking out at least two of antiprotease PR3, antiprotease (MPO) and glomerular basement membrane (GBM) on a nitrocellulose membrane or a nylon membrane. The kit is provided with the ingenious cut-off control strips, one cut-off control strip can interpret two or more detection strips (antigen strips) simultaneously and the judgment result is simpler and more reliable.

Description

technical field [0001] The invention belongs to the field of biotechnology, and relates to a kit for diagnosing diseases, in particular to a kit for detecting autoantibody spectrum related to vasculitis. Background technique [0002] Systemic vasculitis (systemic vasculitis) is a group of diseases with vascular necrosis and inflammation as the main pathological changes. Common vasculitis includes Wegener's granulomatosis, microscopic vasculitis, pulmonary hemorrhage-nephritic syndrome and nodular polyarteritis etc. These diseases all have a common feature: the disease accumulates large and small arteries and veins in various parts of the body, and can cause complex and variable clinical manifestations due to the type, size, location, course of disease, and pathological changes of the blood vessels involved; but they can all detect characteristic Anti-neutrophil cytoplasmic antibody (ANCA) is an important basis for the diagnosis of systemic vasculitis. At the same time, ANC...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/564G01N33/558G01N33/543
Inventor 李想周帅何涛涛陈洪郑丽陈卫陈川
Owner SICHUAN XINCHENG BIOLOGICAL CO LTD
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