Human immunodeficiency virus antibody chemiluminescence immune analyzing diagnose reagent box and method of producing the same

A technology for human immunodeficiency and chemiluminescence immunity, which is applied in the field of immunoanalysis medicine, can solve the problems of uniformity and poor reproducibility of diagnostic kits, and achieve the effects of low cost, strong specificity and affinity, and improved sensitivity

Active Publication Date: 2011-05-18
CHEMCLIN DIAGNOSTICS CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Specifically, the present invention solves the problems of poor uniformity and reproducibility of diagnostic kits in the prior art

Method used

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  • Human immunodeficiency virus antibody chemiluminescence immune analyzing diagnose reagent box and method of producing the same
  • Human immunodeficiency virus antibody chemiluminescence immune analyzing diagnose reagent box and method of producing the same
  • Human immunodeficiency virus antibody chemiluminescence immune analyzing diagnose reagent box and method of producing the same

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] Embodiment 1 prepares HIV antibody diagnostic kit of the present invention

[0035] HIV antibody diagnostic kit of the present invention comprises:

[0036] 1) 96-well or 48-well luminescent microplates pre-coated with anti-FITC antibody;

[0037] 2) FITC-labeled gp41, gp120, gp36 HIV recombinant antigens;

[0038] 3) HIV recombinant antigen labeled with horseradish peroxidase;

[0039] 4) The negative control solution is normal human serum;

[0040] 5) The positive control solution is a positive mixed slurry diluted with newborn bovine serum;

[0041] 6) The sample diluent is 0.1M Tris-HCl buffer, 0.1% BSA, and 0.1% Proclin300, pH7.4;

[0042] 7) The 20-fold concentrated washing solution is 2M Tris-HCl buffer solution containing 10% physiological saline, pH=8.5;

[0043] 8) The chemiluminescent substrates A and B are luminol solution, hydrogen peroxide and special effect enhancer (p-iodophenol) solution respectively.

[0044] Prepare the above kit by the following ...

Embodiment 2

[0067] Embodiment 2 prepares HIV antibody diagnostic kit of the present invention

[0068] Except that plastic beads are used as carrier, HIV recombinant antigen is labeled with alkaline phosphatase, AMPPD is used as chemiluminescent substrate, and the pH of the citrate buffer solution used in the coating process is 4.5, with the same method as in Example 1 Methods Prepare the kit of the present invention.

Embodiment 3

[0069] Embodiment 3 The using method of kit of the present invention:

[0070] 1. Take out the kit from the refrigerator at 4°C, equilibrate at room temperature for 20 minutes, take a bottle of concentrated lotion, add distilled water (1ml+19ml) for later use.

[0071] 2. Take the luminescent plate out of the sealed bag, add 100 μL FITC-labeled HIV recombinant antigen solution (1:2000) to the microtiter plate coated with anti-FITC antibody, and incubate at 37°C for 1 hour.

[0072] 3. Discard the liquid in each well, fill each well with the washing solution, let it stand for 10-20 seconds, shake off the washing solution; repeat the plate washing, and finally pat dry on clean absorbent paper.

[0073] 4. Set a blank control well without adding sample solution, set negative control 3 wells, and positive control 2 wells, take negative control solution, positive control solution and sample solution 50 μl each in the corresponding plate well, and then add 50 μl of enzyme marker , ...

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Abstract

The invention relates to the medical field of immunoassay, which particularly provides a human immunodeficiency virus antibody chemiluminescence immunoassay diagnostic kit and a preparation method thereof. The kit of the invention includes: a solid-phase carrier which is coated by an anti-FITC antibody, FITC-labeled HIV recombinant antigen, enzyme-labeled HIV antigen, chemiluminescence substrate which is acted by the enzyme, negative and positive control liquids. The preparation method of the kit includes the following steps: (1) the solid-phase carrier is coated by the anti-FITC antibody; (2) the FITC-labeled HIV recombinant antigen is obtained by using the FITC-labeled HIV recombinant antigen; (3) the enzyme-labeled HIV antigen is obtained by using the enzyme-labeled HIV antigen; (4) packaging and assembly are carried out. The kit of the invention has low cost and high sensitivity; compared with the existing chemiluminescence immunoassay detection methods, the invention can improve the sensitivity and reproducibility of the analysis and reduce the using amount of the recombinant antigen.

Description

technical field [0001] The invention relates to the field of immunoanalysis medicine, in particular, the invention provides a human immunodeficiency virus (HIV) antibody chemiluminescent immunoassay diagnostic kit and a preparation method thereof. Background technique [0002] Acquired immunodeficiency syndrome, also known as AIDS (AIDS), is an immunodeficiency disease caused by human immunodeficiency virus (HIV) infection. As the disease currently has no effective treatment and preventive vaccines, it is currently spreading at a rate of about 14,000 people being infected. It is estimated that there are nearly 1 million HIV-infected people in my country. If active and effective control measures are not taken, the number of HIV-infected people in my country will reach 10 million by 2010, so the situation is very serious. The diagnosis of HIV infection and the identification of early-infected people are the primary tasks of AIDS prevention and control. Therefore, it is very ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/569G01N33/543G01N21/76
Inventor 应希堂詹先发胡国茂
Owner CHEMCLIN DIAGNOSTICS CO LTD
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