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Multiple index quick detecting method for human immune defect virus antibody

A technology for human immunodeficiency and detection methods, which is applied in the field of multi-index rapid detection of human immunodeficiency virus antibodies, can solve the problems of increased false positive rate, single detection index, low sensitivity and accuracy, and achieves simple operation and accurate detection results. Effect

Inactive Publication Date: 2006-11-08
WUHAN UNIV
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  • Abstract
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  • Application Information

AI Technical Summary

Problems solved by technology

However, the existing HIV rapid detection technology (mainly referring to immunofiltration / chromatography technology) has relatively single detection indicators, and there are problems such as low sensitivity and accuracy.
ELISA has improved sensitivity due to the use of a variety of HIV antigen mixtures, but due to various reasons, its false positive rate has also increased

Method used

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  • Multiple index quick detecting method for human immune defect virus antibody
  • Multiple index quick detecting method for human immune defect virus antibody
  • Multiple index quick detecting method for human immune defect virus antibody

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Experimental program
Comparison scheme
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Embodiment Construction

[0037] 1 Materials and methods

[0038] 1.1 Materials

[0039] 1.1.1 Strains and plasmids Escherichia coli (E.coli) DH5α (cloning host), BL21 (DE3) (expression host) and pNL4-3 (including the full genome sequence of HIV-1), and the expression vector pET28-a are all from this Laboratory preservation.

[0040] 1.1.2 Primers and tool enzymes Primers were synthesized by Shanghai Bioengineering Company. Taq enzyme was purchased from Promega Company. Restriction endonuclease, T4 ligase was purchased from Huamei Bioengineering Company.

[0041] 1.1.3 Main reagents and materials Staphylococcal protein A (SPA) and anti-SPA antibody were purchased from Beijing Benyuan Zhengyang Gene Technology Co., Ltd. Nitrocellulose membrane was purchased from Wuhan Life Science and Technology Co., Ltd. Chloroauric acid was purchased from Shanghai Reagent No. 1 Factory.

[0042] 1.2 Method

[0043] 1.2.1 Construction of HIV antigen expression vector

[0044] Gene manipulation reference literat...

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Abstract

This invention discloses a multi-target quick test method for human immunodeficiency virus antibodies including: 1, expressing five kinds of HIV antigen white dribs and drabs-p24, gp41, gp36, gp120V3, gp120C in a colibacillus by a gene engineering technology, 2, fixing the five antigen whites on a pyroxylin film, 3, dripping armed serum on it and the virus antibody is combined with the antigen by a immunoreaction then adding the A(SPA) labeled by nm gold, 4, cleaning it after it penetrates the film, 5, adding SPA antibodies to increase the amplification to form red spots seen by eyes.

Description

technical field [0001] The invention relates to a detection technology for human immunodeficiency virus antibody, in particular to a multi-index rapid detection method for human immunodeficiency virus antibody. Background technique [0002] Human Immunodeficiency Virus (HIV, commonly known as HIV) is the pathogen that causes Acquired Immunodeficiency Syndrome (AIDS, commonly known as AIDS). Viral antigens (p24, gp41) can be detected in the early stage of HIV infection, but the detection time is very short, usually no more than 2 weeks, and the antigen concentration is very low, making diagnosis difficult. Therefore, most HIV serodiagnosis relies on the detection of antibodies against HIV antigens, and anti-HIV can be detected about 4 to 6 weeks after infection. Usually antibodies against gag-encoded protein p24 emerge first, while antibodies against env-encoded proteins (gp41, gp120) and pol-encoded proteins (p66, p51, p31) appear slightly later. Western blot analysis of a...

Claims

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Application Information

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IPC IPC(8): G01N33/544G01N21/78G01N33/532
Inventor 王业富翟建新
Owner WUHAN UNIV
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