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Kit for quantitative detection of anti-Jo-1 antibody IgG by using magnetic particle chemiluminescence, and preparation method and detection method thereof

A chemiluminescent, quantitative detection technology, applied in the field of immunological detection, can solve problems such as poor repeatability, long reaction time, and narrow linear range

Inactive Publication Date: 2016-04-06
北京贝尔医疗设备有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] Chinese patent CN103105489A discloses an immunoblotting kit for detecting autoimmune disease antibodies and a preparation method thereof, which relates to a immunoblotting kit for detecting antibodies related to various autoimmune diseases, and solves the problem that currently there is no such kit for various autoimmune diseases Technical deficiencies in the western blot product for physical examination screening: on the nitrocellulose membrane or nylon membrane, it contains: respectively composed of dsDNA, Sm / RNP, CCP, SSA, SSB, GAD, ICA, IA-2A, TG, AMA-M2 A total of 10 kinds of at least two natural or recombinant antigens coated with more than two parallel detection lines, 1 high-concentration quality control band, 1 medium-concentration quality control band and 1 low-concentration quality control band bring
The patent discloses a membrane strip immunoassay, which can better explain the problems of low sensitivity, narrow linear range, poor repeatability and long reaction time of the membrane strip immunoassay.

Method used

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  • Kit for quantitative detection of anti-Jo-1 antibody IgG by using magnetic particle chemiluminescence, and preparation method and detection method thereof
  • Kit for quantitative detection of anti-Jo-1 antibody IgG by using magnetic particle chemiluminescence, and preparation method and detection method thereof
  • Kit for quantitative detection of anti-Jo-1 antibody IgG by using magnetic particle chemiluminescence, and preparation method and detection method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0066] A kit for the quantitative detection of anti-Jo-1 antibody IgG using magnetic particle chemiluminescence. The kit includes: Anti-Jo-1IgG calibrator, Anti-Jo-1IgG reagent No. 1, Anti-Jo-1IgG reagent No. 2, Anti-Jo-1IgG magnetic separation reagent, Anti-Jo-1IgG quality control material and cleaning solution.

[0067] Evaluation of analytical performance of the kit of the present invention:

[0068] 1) Accuracy evaluation

[0069] Add the anti-Jo-1 antibody IgG sample A with a concentration of about 200RU / mL and allow its concentration deviation of ±20% to the serum or other corresponding matrix sample B. The volume of A added does not exceed the total volume of A+B 10%, the recovery rate R is calculated according to formula (1), the recovery rate of this method is in the range of 85-115%, see Table 1 for data.

[0070] R = C × ( V 0 + V ) - C 0 × V 0 V × C S × 100 % ... ( 1 )

[0071] R: recovery rate;

[...

Embodiment 2

[0112] A method for preparing a kit for quantitatively detecting anti-Jo-1 antibody IgG using magnetic particle chemiluminescence, including the following steps:

[0113] Step 1. The preparation of the Anti-Jo-1IgG calibrator includes: Anti-Jo-1IgG calibrator diluent preparation steps and Anti-Jo-1IgG calibrator preparation steps:

[0114] The preparation steps of the Anti-Jo-1IgG calibrator diluent include:

[0115] Add 800mL purified water and 11.2g of Tris to the container, stir and mix well, then add 8.5g of sodium chloride and 2mL of Proclin300, stir until completely dissolved, adjust the pH to 7.0-7.5, add 40g of bovine serum white Put the protein into the container, stir until it is completely dissolved, then adjust the pH to 7.0-7.5, dilute to 1L with purified water, filter with a 0.2μm filter, and store the obtained calibrator dilution at 2-8℃ for later use;

[0116] The preparation steps of the Anti-Jo-1IgG calibrator include:

[0117] Prepare the anti-Jo-1 antibody IgG to 0...

Embodiment 3

[0134] On the basis of Example 2, the kit further includes a cleaning solution, and the preparation steps of the cleaning solution:

[0135] Add 800mL purified water, Tris12.1g and NaCl8.5g to a 1L container, stir well until completely dissolved; then add 5g Tween-20 and 5g Triton100, stir well until completely mixed, adjust the pH value to 7.5 -8.0, a constant volume of 1L, filter with a 0.2μm filter, store at a temperature of 2-8°C, dilute and mix the cleaning solution at a ratio of 7.5mL to 100mL with purified water before use.

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Abstract

The invention relates to the technical field of immunology detection, and particularly relates to a kit for quantitative detection of an anti-Jo-1 antibody IgG by using magnetic particle chemiluminescence, and a preparation method and a detection method thereof. The kit for quantitative detection of the anti-Jo-1 antibody IgG by using magnetic particle chemiluminescence includes an Anti-Jo-1 IgG calibrator, an Anti-Jo-1 IgG reagent No. 1, an Anti-Jo-1 IgG reagent No. 2, an Anti-Jo-1 IgG magnetic separation reagent, an Anti-Jo-1 IgG control material and a cleaning solution. The invention also discloses a preparation method and a detection method of the above kit. Based on the traditional film strip immunoassay and enzyme-linked immunosorbent assay, the detection method increases the sensitivity and linearity range by 3-5 orders of magnitude, achieves the quantitative detection, and has the advantages of high sensitivity and specificity, good accuracy, low cost, simple operation and the objective judgment of results. The method cooperates with an automated chemiluminescence immunoassay analyzer to reach complete automation usage, and has broad application prospects.

Description

Technical field [0001] The invention relates to the technical field of immunological detection, in particular to a kit for quantitatively detecting anti-Jo-1 antibody IgG using magnetic particle chemiluminescence and a preparation method and detection method thereof. Background technique [0002] In 1980, it was first reported that anti-Jo-1 antibody was found in the serum of patients with polymyositis (PM). Its antigen is histidyl tRNA synthetase, which is present in the cytoplasm. It was named after the patient’s name as John. It is an anti-Jo-1 antibody. [0003] A large number of clinical studies have confirmed that Anti-Jo-1 antibody (Anti-Jo-1 antibody) is the most important antibody in Myositis-specific antibodies (MSA), which is effective against polymyositis (PM) / cutaneous muscle The diagnosis, differential diagnosis, classification, treatment selection and prognostic evaluation of inflammatory disease (Dermatomyositis, DM) have important clinical significance. It was co...

Claims

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Application Information

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IPC IPC(8): G01N21/76G01N33/553G01N33/531
CPCG01N21/76G01N33/531G01N33/553
Inventor 不公告发明人
Owner 北京贝尔医疗设备有限公司
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