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Test card for rapid detection of epidemic hemorrhagic fever virus antibody, and application of test card

An epidemic hemorrhagic fever, detection and testing technology, applied in the field of immune detection, can solve problems such as inconvenience and time-consuming, achieve high accuracy, avoid false negative results, and facilitate field detection.

Active Publication Date: 2018-11-30
苏州华益美生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the inventors found that although ELISA has good accuracy, it is not simple enough and takes a long time

Method used

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  • Test card for rapid detection of epidemic hemorrhagic fever virus antibody, and application of test card
  • Test card for rapid detection of epidemic hemorrhagic fever virus antibody, and application of test card
  • Test card for rapid detection of epidemic hemorrhagic fever virus antibody, and application of test card

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] The determination of embodiment 1 epidemic hemorrhagic fever virus antigen, anti-human IgG antibody and anti-human IgM antibody

[0030]According to the design of the inventor, the antigen of the amino acid sequence shown in SEQID NO: 1 has been designed from a large amount of antigens of epidemic hemorrhagic fever (EHF) virus as the EHF virus detection antigen; in addition, the inventor has found that commercially available Anti-human antibodies are prone to certain cross-reactions with the above-mentioned detection antigens, resulting in a decrease in detection accuracy. Therefore, two antigens (whose amino acid sequences are shown in SEQ ID NO: 2 and 3 respectively) have been screened out from a large number of human IgG and IgM. Shown), effective anti-human IgG antibody and anti-human IgM antibody can be prepared without complete human IgG and IgM respectively, entrusted Beijing Zhongshan Jinqiao Biotechnology Co., Ltd. to prepare the amino acid sequence shown in SEQ...

Embodiment 2

[0031] Embodiment 2 anti-human IgG, anti-human IgM monoclonal antibody draws film on nitrocellulose

[0032] The anti-human IgG antibody and the anti-human IgM antibody prepared in Example 1 were used to draw a film on a nitrocellulose membrane (NC membrane) with a nitrocellulose membrane, specifically using a Sartori μs (Sartorius) CN104NC membrane with a pore size of 15 μm and a crawling speed of 150S / 4cm. The specific filming process is as follows:

[0033] 1) Preparation of scratching buffer: prepare 0.01 MPBS (phosphate buffer), pH 7.0, containing 1% sucrose, and filter with 0.22 μm.

[0034] 2) Take the scratching buffer and dilute the anti-human IgG antibody, anti-human IgM antibody and anti-rabbit antibody (Beijing Zhongshan Jinqiao Biotechnology Co., Ltd.) respectively according to a certain concentration, wherein the anti-human IgG antibody is diluted to 2.5mg / ml, and the anti-human IgG antibody The IgM antibody was diluted to 1.0mg / ml, and the anti-rabbit antibody...

Embodiment 3

[0036] The determination of embodiment 3 fluorescent microspheres and the coupling with antigen, antibody

[0037] Take carboxyl group fluorescent microspheres from different sources, dilute them with 0.01M PBS at 1:10; use 0.01M PBS to adjust to zero, and scan in the wavelength range of 400nm to 600nm to see if there is a narrow absorption peak at a wavelength of 480nm (indicating fluorescence Uniform microsphere particle size); Dilute the concentration of fluorescent microspheres to 2mg / ml with 0.01M PBS, spot 10μl on the NC membrane, and check to see if the fluorescence value is ≥10 4 . After testing, the fluorescent microspheres with carboxyl groups purchased from Ocean Nanotech have the best effect, so this fluorescent microspheres were selected. The specific antigen and antibody coupling process is as follows:

[0038] 1) EHF antigen-labeled fluorescent microspheres: Dilute the fluorescent microspheres to 1 mg / ml with activation buffer (0.05M MES (2-(N-morpholino)ethan...

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Abstract

The invention provides a test card for rapid detection of an epidemic hemorrhagic fever virus antibody. The test card comprises a bottom plate, a sample pad, a nitrocellulose membrane, a water absorption pad and a fluorescent microsphere pad, wherein the fluorescent microsphere pad adsorbs fluorescent microspheres marked by epidemic hemorrhagic fever virus antigens and fluorescent microspheres marked by quality control proteins; and an anti-human IgG antibody, an anti-human IgM antibody and an anti-quality control protein antibody are sequentially adsorbed on the nitrocellulose membrane. In addition, the invention provides a preparation method of the test card, an application of the test card to the rapid detection of the epidemic hemorrhagic fever virus human antibody, and the like.

Description

technical field [0001] The invention belongs to the technical field of immune detection, and in particular relates to a test card for rapid detection of antibodies to epidemic hemorrhagic fever virus (including IgG and IgM), an application thereof, and the like. Background technique [0002] Epidemic hemorrhagic fever (EHF) is a natural focal disease caused by a virus. In 1982, the World Health Organization (WHO) named it hemorrhagic fever with renal syndrome (hemorrhagic fever with renal syndromes, HFRS). It is an acute viral infectious disease characterized by fever, bleeding tendency and kidney damage. The disease is mainly distributed in Eurasia, but the spread of HFRS virus is almost all over the world. [0003] The diagnostic method of epidemic hemorrhagic fever is to detect virus antigen, anti-hemorrhagic fever virus IgG and IgM antibodies, viral RNA, etc. from serum, all of which are used as the basis for diagnosis. EHF-IgM antibodies can be detected within 2 to 3...

Claims

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Application Information

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IPC IPC(8): G01N33/68G01N33/569
CPCG01N33/56983G01N33/68
Inventor 李振勇郜迎晨牛莉娜张文艳张鑫
Owner 苏州华益美生物科技有限公司
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