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Colloidal gold semi-quantitative quick immunity diagnosis test-paper stripe

A rapid immunoassay, semi-quantitative technology, applied in biological testing, material inspection products, etc., to achieve the effect of simple production, simple testing procedures, and convenient on-site testing

Inactive Publication Date: 2012-08-22
LANZHOU UNIVERSITY
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  • Colloidal gold semi-quantitative quick immunity diagnosis test-paper stripe

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specific Embodiment 2

[0009] (1) Purchasing type A and III influenza virus-specific antigen neuraminidase (NA) and hemagglutinin protein to immunize mice, and reassured to obtain mouse-derived specific monoclonal antibodies IgG1 and IgG2 against the two antigens. (2) IgG1 was selected as the gold-labeled antibody, and the sodium citrate reduction method was used to prepare and determine that the optimal size of labeled gold nanoparticles was 15 nm, and centrifuged at 7300 rpm / min for 20 minutes to prepare a stock solution. 15nm colloidal gold and labeled monoclonal antibody were used to determine the optimal amount of protein, and the pH9.0 borate buffer was used to make the labeled monoclonal antibody a gradient of 5-50Ug / ml. After adding the gold stock solution, 10% NaCl was added for stability experiments. Minutes later, centrifuge and measure OD580nm, and the protein concentration is 10ug / ml when the optical density is stable. After preparation, add PEG to make colloidal gold stable solution. ...

specific Embodiment 3

[0011] (1) Purchasing pure staphylococcal protein A (SPA) to determine the protein content (2) Using the sodium citrate reduction method to determine the size of the optimally labeled gold nanoparticle for the SPA used, centrifuged at 2000rpm / min for 30 minutes to prepare a stock solution . Use 40nm colloidal gold and SPA to determine the optimal amount of protein, use PH9.0 borate buffer to make the labeled SPA a gradient of 5-50Ug / ml, add 10% NaCl after adding gold stock solution for stability experiment, and centrifuge for 5 minutes The OD580nm was measured, and the SPA protein concentration was 4.6ug / ml when the optical density was stable. After preparation, add PEG to make colloidal gold stable solution. (3) Absorb the marked gold standard SPA with a sterile absorbent sponge with a width of 1 cm, a width of 1.5 cm, and a thickness of 0.08 cm to make a gold marker pad. In a dark environment at 4°C, blow dry slowly with nitrogen gas for use. (4) Select sterile cellulose a...

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Abstract

The present invention relates to a colloidal gold semiquantitative immunologic diagnosis test paper preparing and assembly method. Said test paper utilizes immunity chromatograph principle capable of rapid visual inspection nano colloidal gold marker and sample combined colour, judging detective semiquantitative result through test zone with parallel reference zone colour contrast.

Description

[0001] Technical field The present invention relates to a kind of nano-colloidal gold test paper for rapid semi-quantitative detection of pathogens (antibodies or antigens), and also relates to the design and assembly organization of the test paper. Background technique [0002] Existing test strips for immunodiagnosis are the most convenient and fast among immunodiagnostic reagents. According to diagnostic categories, they can be divided into infectious diseases, endocrine, tumors, drug testing, etc. However, the current test strips are mainly based on the qualitative detection of colloidal gold from the perspective of the methodology of result judgment. The test results can only determine the presence or absence of pathogens but cannot determine whether their content is higher than a certain diagnostic standard. In practical applications, judging the content of a certain substance to be detected is of more reference value for clinical diagnosis. Qualitative colloidal gold im...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/53G01N33/52
Inventor 李红玉张波
Owner LANZHOU UNIVERSITY
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