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HIV (human immunodeficiency virus) antigen antibody detection card and preparation method thereof

A technology of antigen-antibody and detection card, which is applied in the field of HIV antigen-antibody detection card and its preparation, can solve the problems of sensitivity to be improved, large sample consumption, etc., and achieve the effect of shortening the window period, reducing sample consumption, and high affinity

Inactive Publication Date: 2017-05-31
山东康华生物医疗科技股份有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] At present, the existing technology has realized the detection of HIV antigen and antibody, but the sample volume is relatively large, about 60 to 100 μL, and the sensitivity needs to be improved

Method used

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  • HIV (human immunodeficiency virus) antigen antibody detection card and preparation method thereof
  • HIV (human immunodeficiency virus) antigen antibody detection card and preparation method thereof
  • HIV (human immunodeficiency virus) antigen antibody detection card and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0048] Such as figure 1 and figure 2 As shown, a test card for detecting HIV antigen and antibody using a biotin-avidin system includes a card cover 1, a card body 2 and a test strip 3. The card cover 1 is provided with a diluent sample hole 11, a sample sample The sample hole 12, the observation hole 13 and the installation mark hole 14, the inside of the card body 2 is provided with a groove 21 for placing the test strip 3. The sample cushion layer 31 on the test strip 3 corresponds to the diluent sampling hole 11 and the sample sampling hole 12 on the card cover 1; the colloidal gold layer 32 and the nitrocellulose membrane 33 on the test strip 3 correspond to the card cover 1 corresponding to the observation hole 13.

[0049] Such as image 3 and Figure 4 As shown, the test strip 3 is sequentially provided with a sample cushion layer 31, a colloidal gold layer 32, a nitrocellulose membrane 33, and a water-absorbing layer 34. The sample cushion layer 31 is a polyester...

Embodiment 2

[0063] A method for preparing a detection card for detecting HIV antigen and antibody by using a biotin-avidin system, which is assembled from the above-mentioned card cover 1, card body 2 and test strip 3, and the test strip 3 is sequentially provided with a sample cushion 31. Colloidal gold layer 32, nitrocellulose membrane 33 and water-absorbing layer 34.

[0064] 1. The preparation process of the sample cushion layer 31 is as follows:

[0065] (1) Biotin-labeled p24 antibody

[0066] 1.1 Configuration of p24 antibody solution: Dilute p24 antibody to 1mg / mL with 0.1mol / L sodium bicarbonate buffer (pH 8.0), and then fully dilute 1mL p24 antibody with 0.1mol / L sodium bicarbonate buffer (pH 8.0). Dialysis;

[0067] 1.2 Configuration of biotin solution: Dissolve 1 mg of biotin N-hydroxysuccinimide ester (NHSB) in 1 mL of dimethyl sulfoxide (DMSO);

[0068] 1.3 Labeling treatment: add 120 μL NHSB solution (containing 120 μg NHSB) to 1 mL p24 antibody solution, keep stirring a...

Embodiment 3

[0091] Purchase HIV-1p24 antigen national reference product (batch number 220015-20150505) from China Food and Drug Control Institute, a total of 44 copies, including 10 positive, 20 negative and 10 minimum detection amount (sensitivity) reference products (L1~L10 , the antigen concentration is 20, 10, 5, 2.5, 1.25, 0.625, 0.313, 0.156, 0.0781, 0U / mL), 2 antigen precision reference products, 2 antigen antibody precision reference products. According to the operation method of the above-mentioned test card, test the above-mentioned national reference products, the results are as follows:

[0092] Positive reference product coincidence rate: 10 positive reference product test results did not appear negative, and the positive coincidence rate was 10 / 10;

[0093] Negative reference product coincidence rate: 20 negative reference product test results were not positive, and the negative coincidence rate was 20 / 20;

[0094]Minimum detection amount reference product: For the HIV anti...

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Abstract

The invention belongs to the technical field of biological detection, and particularly relates to an HIV (human immunodeficiency virus) antigen antibody detection card and a preparation method thereof. The detection card comprises a card cover, a card body and a test strip, wherein the card cover is sequentially provided with a dilute solution adding hole, a sample adding hole, an observation hole and an installation marking hole; a groove for holding the test strip is arranged in the card body; and the test strip is sequentially provided with a sample pad layer, a colloidal gold layer, a nitrocellulose film and a water-absorbing layer. The preparation method comprises the following steps: preparing colloidal gold; respectively marking recombinant HIV-1 / HIV-2 antigens and mouse anti-p24 monoclonal antibodies with the colloidal gold, marking p24 antibodies with biotin, immobilizing the colloidal gold and biotin markers, coating the nitrocellulose film, preparing the test strip, and buckling the detection card. The detection card provided by the invention can detect HIV antigen antibodies, and is high in sensitivity and low in sample consumption.

Description

technical field [0001] The invention relates to the technical field of biological monitoring, in particular to an HIV antigen antibody detection card and a preparation method thereof. Background technique [0002] AIDS, the full name is "Acquired Immune Deficiency Syndrome" (Acquired Immune Deficiency Syndrome), English abbreviation AIDS, is an infectious disease caused by human immunodeficiency virus (Human immunodeficiency virus, HIV). HIV itself does not cause any disease, but when the immune system is destroyed by HIV, the human body loses resistance and is infected with other diseases leading to death. [0003] HIV infection can be established by detecting HIV-specific antibodies, antigens, nucleic acids, or virus isolation and culture. The standard diagnostic item routinely used is serological antibody detection. From the advent of the first generation of HIV antibody detection reagents in 1985 to the present, HIV serological detection reagents have developed to the f...

Claims

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Application Information

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IPC IPC(8): G01N33/569G01N33/577
CPCG01N33/56988G01N33/577
Inventor 杨致亭隋振国陈利军魏红王明儒杨明霞
Owner 山东康华生物医疗科技股份有限公司
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