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Anti-double-stranded DNA antibody detection kit and application thereof

An antibody detection and kit technology, applied in the field of anti-double-stranded DNA antibody detection kits

Pending Publication Date: 2021-11-19
SICHUAN MACCURA BIOTECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] Based on this, it is necessary to provide an anti-double-stranded DNA antibody detection kit and its application for the false positive problem caused by positively charged substances in the sample in the anti-double-stranded DNA antibody

Method used

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  • Anti-double-stranded DNA antibody detection kit and application thereof
  • Anti-double-stranded DNA antibody detection kit and application thereof
  • Anti-double-stranded DNA antibody detection kit and application thereof

Examples

Experimental program
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Effect test

preparation example Construction

[0063] In another aspect, the present invention provides the preparation method of the anti-double-stranded DNA antibody IgG assay kit as described above, the preparation method comprising: preparation of double-stranded DNA antigen; preparation of solid phase carrier; secondary antibody labeled with signal substance preparation.

[0064] In some embodiments, the solid phase carrier is coupled with streptavidin, the double-stranded DNA is linked with biotin, and the double-stranded DNA is coated on the solid phase by biotin and streptavidin. phase carrier. The preparation method is as follows:

[0065] (1) adding the streptavidin solution into the solid phase carrier to carry out the coupling reaction to obtain the streptavidin coupled solid phase carrier;

[0066] (2) adding the biotinylated double-stranded DNA antigen solution to the streptavidin-coupled solid-phase carrier for incubation and coating to obtain the solid-phase carrier coated with the double-stranded DNA ant...

Embodiment 1

[0083] In this example, concentration 0 (no heparin sodium, control), concentration 1 (0.01 mg / mL), concentration 2 (0.1 mg / mL), concentration 3 (1 mg / mL) of heparin sodium were added to reagent R1 respectively. , Concentration 4 (10mg / mL), Concentration 5 (100mg / mL), the magnetic particle concentration of reagent R1 is 0.4mg / mL, the coating amount of double-stranded DNA is 5μg / mg, and the other ingredients are the same as in Table 1. The test results are shown in Table 2;

[0084] Table 2: Detection results of different heparin sodium concentrations

[0085]

[0086]

[0087] Note: Samples Q1-Q10 were negative by the INOVA (dsDNA) kit and identified as healthy population samples; samples S11-S20 were positive by the INOVA (dsDNA) kit and identified as clinical patient samples;

[0088] Conclusion: As the amount of heparin sodium increases, the RLU value gradually decreases, and the overall trend gradually slows down. When the content of heparin sodium is 1 mg / mL, it re...

Embodiment 2

[0090] Under the condition that the concentration of heparin sodium in reagent R1 was 1 mg / mL and the coating ratio of double-stranded DNA was 5 μg / mg, different concentrations of magnetic particles (0.2 mg / mL, 0.4 mg / mL, 0.6 mg / mL), and the rest of the ingredients are the same as in Table 1. The test results are shown in Table 3;

[0091] Table 3: Detection results of magnetic particles with different concentrations

[0092]

[0093]

[0094] Conclusion: Compared with the kit without adding heparin sodium, after adding heparin in reagent R1, the RLU value detected by the reagent R1 magnetic particle concentration is significantly reduced. It can be considered that adding heparin sodium in reagent R1 can Effectively eliminate false positives.

[0095] Compared with 0.6 mg / mL, when the concentration of magnetic particles was 0.4 mg / mL, the RLU value of sample detection changed more. That is, when the concentration of magnetic particles increases to 0.4 mg / mL, the chan...

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Abstract

The invention discloses an anti-double-stranded DNA antibody detection kit, which comprises an R1 reagent and an R2 reagent, the R1 reagent comprises a solid phase carrier coated with double-stranded DNA, an R1 dispersant and heparin, the R2 reagent comprises a second antibody labeled with a signal substance, the concentration of the heparin in the R1 reagent is 1 mg / mL or more, and the heparin exists in the R1 reagent; or the kit comprises the R1 reagent, the R2 reagent and an R3 reagent, the R1 reagent comprises a solid phase carrier coated with double-stranded DNA and a dispersant R1, the R2 reagent comprises a secondary antibody labeled with a signal substance, the R3 reagent comprises a dispersant R3, heparin exists in the R1 reagent or the R3 reagent or exists in the R1 reagent and the R3 reagent at the same time, and the concentration of heparin in the R1 reagent is preferably 1 mg / mL or above; and / or the concentration of heparin in the R3 reagent is preferably more than 1 mg / mL; and the second antibody is an antibody for resisting species immune globulin from the tested sample. The invention also discloses application of the kit in preparation of a diagnostic reagent or kit for diagnosing systemic lupus erythematosus.

Description

technical field [0001] The invention relates to the technical field of in vitro detection, in particular to an anti-double-stranded DNA antibody detection kit and its application. Background technique [0002] In the IVD industry, what we usually refer to as an immune response refers to the specific binding of an antigen to the corresponding antibody induced during the immune response. By using a known antigen to detect the corresponding antibody or a known antibody to detect the corresponding antigen, the purpose of qualitative or quantitative determination of a certain antigen or antibody in the test sample is achieved. [0003] Currently, commonly used detection methods for antigen-antibody reactions include fluorescence immunoassay, enzyme-linked immunoassay, and chemiluminescence. Among them, the chemiluminescence method generally immobilizes a certain known antigen or antibody on a specific solid-phase carrier, and then adds the sample to be tested, and detects the su...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68G01N33/543G01N33/53
CPCG01N33/6854G01N33/54326G01N33/5308G01N2800/104
Inventor 王小波蒋小龙
Owner SICHUAN MACCURA BIOTECH CO LTD
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