Kit for detecting anti-double-stranded DNA antibody, composition and application

Technology of a kit and composition

Active Publication Date: 2020-08-18
SICHUAN MACCURA BIOTECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

But in this way, the advantages of acridine chemiluminescence, such as high luminous efficiency, easy operation, high sensitivity and wide linear range, cannot be utilized.

Method used

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  • Kit for detecting anti-double-stranded DNA antibody, composition and application
  • Kit for detecting anti-double-stranded DNA antibody, composition and application
  • Kit for detecting anti-double-stranded DNA antibody, composition and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0087] The preparation of embodiment 1 reaction reagent

[0088] R1：

[0089] 1. Take the double-stranded DNA antigen (purchased from Sichuan Mike Bio-New Material Technology Co., Ltd., hereinafter abbreviated as Sichuan New Material), add it to the magnetic particles (purchased from Sichuan New Material) and mix well according to the ratio of double-stranded DNA to magnetic particles: Add 1-10ug / mg, place on a roller mixer and mix at room temperature for 2 hours;

[0090] 2. After the reaction is completed, place the magnetic particles coated with double-stranded DNA on a magnetic separator for 2 minutes; discard the supernatant after clarification with liquid, add buffer to resuspend the magnetic particles, and mix with a roller mixer at room temperature Reaction for 0.5 hours;

[0091] 3. Take the coated immunomagnetic particles and place them on the magnetic separator to stand still. After the liquid is clarified, discard the supernatant; add the buffer to resuspend the...

Embodiment 2

[0105] Embodiment 2 detection method

[0106] Use Mike Biological i3000 automatic chemiluminescence instrument for the following detection;

[0107] 1. Mix the sample (10 μL) with R3 (100 μL) and R1 (50 μL) in Example 1, and react at 37° C. for 10 minutes;

[0108] 2. After washing, add R2 (100 μL) and react at 37°C for 10 minutes;

[0109] 3. After washing again, add hydrogen peroxide solution and alkali solution to produce chemiluminescent reaction, and measure the relative luminescence value (RLU).

Embodiment 3

[0110] Example 3 Evaluating the Effect of Acridone on Improving the Coefficient of Variation

[0111] According to the method in Example 2, the anti-double-stranded DNA antibody in the serum of a patient clinically diagnosed as systemic lupus erythematosus was used as the detection object, and the coefficient of variation (CV) of RLU after repeated detection of the same sample was investigated.

[0112] Wherein, R1, R2 and R3 prepared in Example 1 were used in the control group A. The experimental group added acridone, and the rest were the same as the control group A. Specifically, 0.01 mg / L acridone was added to R2 and R3 of experimental group A1 respectively; 0.02 mg / L acridone was added to R2 and R3 of experimental group A2 respectively; Add the acridone of 0.10mg / L; Add the acridone of 0.50mg / L respectively in R2 and R3 of experimental group A4; Add the acridone of 1.00mg / L respectively in the R2 and R3 of experimental group A5, all the other with The control group was th...

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Abstract

The invention relates to a kit for detecting an anti-double-stranded DNA antibody. The kit comprises a solid-phase support coated with a second component, and a first component marked with an acridinechemiluminiscence substance. The kit is characterized in that the kit further comprises an acridine non-chemiluminiscence substance, and under the alkaline condition, the ninth position of the acridine non-chemiluminiscence substance cannot form a four-membered ring with peroxide. The kit can improve the relative variation coefficient of the detection result of the anti-double-stranded DNA antibody. The invention also relates to a corresponding composition and use for detecting anti-double-stranded DNA antibodies.

Description

technical field [0001] The present invention relates to the detection of anti-double-strand DNA antibodies, in particular to the detection of double-strand DNA antibodies based on chemiluminescence. Background technique [0002] Anti-double-stranded DNA antibodies are antibodies that recognize natural double-stranded DNA. On the one hand, anti-double-stranded DNA antibodies can be used as highly specific antibodies for systemic lupus erythematosus (SLE), which have a higher positive rate in active SLE and a lower positive rate in inactive SLE. However, it should be noted that a negative anti-double-stranded DNA antibody test does not necessarily rule out SLE, and a comprehensive evaluation should be combined with other indicators. On the other hand, it is reported that double-stranded DNA antibodies are also associated with Sjogren's syndrome, drug-induced lupus, mixed connective tissue disease, kidney damage, and vasculitis. The detection results of anti-double-stranded D...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K16/44G01N33/53G01N21/76
CPCC07K16/44G01N21/76G01N33/53
Inventor 王小波田君喜龙腾镶
Owner SICHUAN MACCURA BIOTECH CO LTD
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