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Anti-cardiolipin antibody detection kit and use thereof

A technology for detecting kits and cardiolipin antibodies, applied in biological testing, measuring devices, material inspection products, etc., to achieve good stability, high specificity and sensitivity

Active Publication Date: 2009-08-05
上海尧浩生物技术有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the existing methods for detecting anticardiolipin antibodies include gold standard and chemiluminescence detection antibodies, which can only be used for simple qualitative analysis, while chemiluminescence methods require special equipment, and most of the free antibodies in serum are detected by enzyme-linked methods. Immunosorbent assay (ELISA), which has high specificity and sensitivity and does not require special instruments, can be qualitative and quantitative, can detect antibody classes and subclasses, is easy to standardize, and has the advantages of simple operation, correct results, and low cost. It has been widely adopted by hospitals and has become the worldwide standard for quantitative determination of ACA.

Method used

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  • Anti-cardiolipin antibody detection kit and use thereof
  • Anti-cardiolipin antibody detection kit and use thereof
  • Anti-cardiolipin antibody detection kit and use thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0055] The preparation of embodiment 1 kit

[0056] 1. Cardiolipin antigen (source: Sigma Company), compared by thin-layer chromatography and high-performance liquid chromatography, its RF value and peak shape are consistent with standard cardiolipin, with a purity of >98%.

[0057] Storage of cardiolipin antigen: Store the cardiolipin antigen whose purity has been identified and meets the standard at -20°C for later use.

[0058] 2. Preparation of pre-coated enzyme-linked plates coated with cardiolipin antigen:

[0059] 2.1 Coating: Add the cardiolipin antigen prepared in step 1 to carbonate buffer (pH9.6, Na 2 CO 3 1.6g / L, NaHCO 3 h 2 (203.2g / L formula), mix well, add in the well of enzyme-linked plate, 100 microliters in every well, hatch overnight, with the phosphate buffer saline (20X, Nacl 85g / L, T-20 0.5ml) containing Tween / L, formula) wash the enzyme-linked plate, and complete the preparation of the pre-coated enzyme-linked plate coated with ACA antigen;

[0060...

Embodiment 2

[0089] Qualitative detection of embodiment 2 ACA

[0090] Adopt the following method to use the kit prepared by embodiment 1:

[0091] a. Adding samples: Take the pre-coated enzyme-linked plate coated with ACA antigen, set blank control wells and negative serum control wells for each experiment, add 100 microliters of sample diluent to each well, and then add negative serum control in turn , positive serum control well, and 100 microliters each of the sample, mix well, stick a self-adhesive strip, and incubate at 37°C for 30 minutes;

[0092] b. Plate washing: Discard the liquid in each well, dilute the washing liquid 20 times with distilled water, fill each well, let it stand for 30 seconds, shake off the washing liquid, wash the plate repeatedly, and finally pat dry;

[0093] c. Add enzyme: add 100 microliters of IgG enzyme-labeled antibody diluted with enzyme-labeled antibody diluent, do not add to the blank control well, stick a self-adhesive strip, and incubate at 37°C f...

Embodiment 3

[0104] Embodiment 3 kit stability test

[0105] For the stability test of the kit, five batches of different batches were selected, and 2 boxes in each batch were used for the stability test. Two batches of kits were placed at 37°C for 3 days, and three batches were placed at 4°C for 6 months. Keep away from light

[0106] Table 1. Test results of physical properties of the kit

[0107]

[0108]

[0109] Table 2. Test results of main performance indicators

[0110]

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Abstract

The invention relates to an anti-cardiolipin antibody detecting reagent kit and the application thereof. The anti-cardiolipin antibody detecting reagent kit comprises a pre-coated enzyme-linked plate which is coated by ACA antigen, IgG enzyme label antibody solution which is diluted by enzyme label antibody diluent, scrub solution, sample diluents, negative contrast solution, positive contrast solution, visualization solution and stop solution. The reagent kit has high specificity and sensitivity and good reagent stability and provides a potent and reliable detection basis for clinical diagnosis.

Description

technical field [0001] The invention relates to an anticardiolipin antibody detection kit and application thereof. Background technique [0002] Anticardiolipin antibodies (ACL) are autoantibodies that target negatively charged cardiolipin on platelet and endothelial cell membranes. Common in systemic lupus erythematosus (SLE) and other autoimmune diseases. The antibody is closely related to thrombosis, platelets, spontaneous abortion or intrauterine stillbirth. Studies have confirmed that many factors are closely related to the occurrence of ACA, and the common reasons are: [0003] (1) Autoimmune diseases: such as systemic lupus erythematosus (SLE), scleroderma of rheumatoid arthritis (RA), etc.; [0004] (2) Viral infection: such as adenovirus, rubella virus, varicella virus, mumps virus and other infections; [0005] (3) Other diseases: such as mycoplasma system diseases, etc.; [0006] (4) Oral certain drugs: such as chlorpromazine, phenothiazine, etc.; [0007] C...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/53G01N33/543G01N21/31
Inventor 崔贻芬
Owner 上海尧浩生物技术有限公司