Kit for detecting anti-double-stranded DNA antibody, composition and application
Technology of a kit and composition
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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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