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Preparation method of membrane strip of kit for detecting autoantibody spectrum related to autoimmune liver disease (AILD) and kit composed of same

An autoantibody and autoimmune technology, applied in the biological field, can solve the problems of not being able to be used as a basis for diagnosis, easy to misjudgment and omission, complicated operation, etc., and achieve the effect of simple and reliable result judgment and uniform width.

Active Publication Date: 2013-02-20
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. At present, there is no improved Western blot method that can simultaneously detect 7 autoantibodies related to autoimmune liver disease
[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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  • Preparation method of membrane strip of kit for detecting autoantibody spectrum related to autoimmune liver disease (AILD) and kit composed of same
  • Preparation method of membrane strip of kit for detecting autoantibody spectrum related to autoimmune liver disease (AILD) and kit composed of same
  • Preparation method of membrane strip of kit for detecting autoantibody spectrum related to autoimmune liver disease (AILD) and kit composed of same

Examples

Experimental program
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Effect test

Embodiment 1

[0034] see Figure 1 to Figure 2 as shown, figure 2 The CO quality control line is the critical quality control zone; the kit for detecting autoimmune liver disease-related autoantibody spectrum in this embodiment includes membrane strips, enzyme labeling solution, substrate and concentrated washing incubation solution, wherein enzyme labeling solution, bottom The selection of substances and concentrated washing incubation solution all belong to the prior art, and for enzyme labeling solution, substrate and concentrated washing incubation solution, selecting products of the prior art can also realize the technical scheme of the present invention.

[0035] The membrane strip is composed of slides and antigen strips, critical quality control strips, and functional quality control lines fixed on the slides in sequence. The antigen strips are composed of AMA-M2, LKM-1, LC-1, SLA, F-actin, gp210 At least two of Sp100 and Sp100 are independently streaked onto a nitrocellulose memb...

Embodiment 2

[0044] This example is carried out on the antigen bands formed on nitrocellulose membrane or nylon membrane by 7 independent lines of AMA-M2, LKM-1, LC-1, SLA, F-actin, gp210 and Sp100 test determination. 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 122 fresh seru...

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 detect random negative samples with this kit, and scan the gray scale, and calculate the mean...

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Abstract

The invention relates to a preparation method of a membrane strip of a kit for detecting autoantibody spectrum related to an AILD and the kit composed of the same. According to the preparation method of the membrane strip and the kit, the membrane strip is composed of a carrier piece and an antigen band, a critical quality control band and a functional quality control line which are sequentially fixed on the carrier piece. The antigen band is formed by at least two of AMA-M2, LKM-1, LC-1, SLA, F-actin, gp210 and Sp100 through independent marking to a nitrocellulose membrane or a nylon membrane. The ingenious critical quality control band is provided, one critical quality control band can play a role in interpreting two or even more detection bands (antigen bands), and result determination 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 method for preparing membrane strips of a kit for detecting autoimmune liver disease-related autoantibody spectrum and the kit thereof. Background technique [0002] Autoimmune liver disease is a kind of chronic progressive liver disease of unknown etiology, which is closely related to autoimmune reaction. Its early clinical manifestations are hidden and difficult to distinguish from chronic viral hepatitis. The treatment methods of these two diseases are quite different. If not treated properly, it can eventually develop into liver fibrosis and cirrhosis, and the only way to prolong the patient's life is through liver transplantation. Therefore, early diagnosis and differential diagnosis of autoimmune liver disease is very urgent and necessary. Autoimmune liver diseases have specific antibodies, which are an important basis for early ...

Claims

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Application Information

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IPC IPC(8): G01N33/576G01N33/564G01N33/531G01N33/558
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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