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Lupus erythematosus detection protein chip and kit thereof

A lupus erythematosus and protein chip technology, which is applied in the field of protein chip disease detection products, can solve the problems of lack of multi-index protein detection products, lack of effectiveness and pertinence, and failure to display index detection values, so as to reduce detection costs and improve detection indicators. Appropriateness, the effect of improving the efficiency of detection

Inactive Publication Date: 2010-06-09
上海裕隆生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although the automated immune analyzer can detect multiple ANA and ENA markers at the same time, many indicators in the test results are not what doctors need to refer to, which wastes time and cost; there are even some indicators that cannot be detected by this method
The ELISA detection method can only be an overall census of ANA and ENA, and cannot display the detection value of specific indicators; or carry out targeted single-indicator detection, but only one indicator can be detected at a time
If you want to check the expression levels of all markers related to SLE by ELISA, you must perform multiple tests, which is time-consuming and requires a large number of serum samples from the patient
It can be seen that these tests in the domestic market are obviously lacking in effectiveness and pertinence, and there is a lack of multi-index protein detection products specifically for these latest research results.

Method used

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  • Lupus erythematosus detection protein chip and kit thereof
  • Lupus erythematosus detection protein chip and kit thereof
  • Lupus erythematosus detection protein chip and kit thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] Preparation of a protein chip for detecting lupus erythematosus

[0030] The main steps are: (1) Prepare the matrix nitrocellulose membrane; (2) Design the chip, determine the arrangement of the lattice and the spotting position; (3) Draw the antibody from the 384-well plate and spray it on the nitrocellulose (4) assemble a nitrocellulose membrane, and carry out sealing, drying, packaging, and preservation.

[0031] The preparation method of the above-mentioned protein chip for detecting multiple tumor marker antibodies in males specifically includes the following steps:

[0032] 1. Determination of antigen and antibody detection of lupus erythematosus markers

[0033] Determination of detection index antigen:

[0034] Lupus erythematosus markers are all purified antigens extracted from human serum.

[0035] Determination of detection index antibodies:

[0036] The detection antibody pairs used are all mouse hybridoma monoclonal antibody pairs, and their detection p...

Embodiment 2

[0042] Kit preparation

[0043] A lupus erythematosus detection protein chip kit, composed of lupus erythematosus detection protein chip, standard substance, quality control substance, sample diluent, enzyme label working solution, concentrated washing solution, detection solution; each part is bottled, with paper template fixed in the box (such as figure 2 ), not limited to the placement method, it is used together with the test. Here's how the parts are prepared:

[0044] 1. The Lupus Erythematosus Detection Protein Chip is a 6×8-well plate, with nitrocellulose membrane as the base, and the assembly method is shown in ZL032298900. After the completion of the appearance, see figure 1 .

[0045] 2. The standard product provides 6 bottles, each bottle is 0.25ml.

[0046] With 3% BSA and 0.05% preservatives (Proclin TM 300) of phosphate buffer (pH 7.4) configuration, wherein the final anti-dsDNA, anti-ssDNA, anti-Histon, anti-SS-A (Ro60), anti-SS-B (La), anti-Sm and anti-...

Embodiment 3

[0064] Application and detection of protein chips and kits

[0065] The application of the protein chip for detecting lupus erythematosus according to the present invention mainly adopts the antigen-antibody-enzyme-labeled secondary antibody method to detect the lupus erythematosus marker antibody in human blood. The antibodies were respectively bound to the corresponding antigens immobilized on the chip. After drying, the enzyme-labeled secondary antibody was added to react, and the unbound secondary antibody was washed away. After 1 minute after adding the detection solution, the signal was scanned and collected. The signal intensity of the sample point It is directly proportional to the concentration of antibody in serum.

[0066] In the above-mentioned application of the protein chip kit for detecting lupus erythematosus, the specific methods for its antigen-antibody reaction and detection are as follows:

[0067] (1) The chips sprayed with antigen, positive control and n...

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Abstract

The invention discloses lupus erythematosus detection protein chip and kit thereof. The chip comprises a substrate, protein markers distributed in an array type and control point coatings, wherein the markers and the control point coatings are seven antigen markers, positive controls and negative controls comprising dsDNA (double-stranded deoxyribonucleic acid), ssDNA (single stranded deoxyribonucleic acid), Histon, SS-A(Ro60), SS-B(La), Sm (smith) and RNP (ribonucleoprotein) / Sm which are uniformly distributed on the substrate in a dot matrix mode. The invention has comprehensive and appropriate detection indexes and consistent applied reaction conditions, can detect a plurality of indexes at a time, is convenient and quick, greatly improves the detection efficiency and reduces the detection cost. The invention can be suitable for aided diagnosis of suspected lupus erythematosus disease people.

Description

technical field [0001] The invention relates to a protein chip disease detection product, in particular to a lupus erythematosus detection protein chip and a kit thereof. Background technique [0002] Systemic Lupus erythematosus (SLE) is an autoimmune disease involving multiple factors that can involve multiple systems and organs throughout the body, accompanied by a variety of immunological abnormalities, and can cause renal failure, infection, death due to damage to the central nervous system. If there is multi-system damage, the diagnosis is not difficult, but at this time, most of the diseases are in the middle and late stages, and the patients have already involved important organs, and the prognosis is poor. In the early stage of SLE, only one or two systems may be involved, and it is easy to be misdiagnosed as other diseases. Therefore, early diagnosis of SLE is particularly important to improve its prognosis and prevent vital organ involvement. [0003] The most ...

Claims

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Application Information

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IPC IPC(8): G01N33/53
Inventor 穆海东汪宁梅穆宇豪刘纲王通
Owner 上海裕隆生物科技有限公司
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