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Time resolved immunofluorescence detection kit for rabies virus nucleoprotein, and preparation method thereof

A technique for immunofluorescence detection and detection kit, which is applied in the field of virus protein immunoassay, and achieves the effects of simple production, high detection sensitivity and short operation process.

Active Publication Date: 2013-09-25
GUANGZHOU PROMISE BIOLOGICAL PROD
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

In traditional fluorescent immunoassays, the problem of high background has not been well resolved
There is no prior art report on quantitative detection of rabies virus nucleoprotein by TRFIA method

Method used

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  • Time resolved immunofluorescence detection kit for rabies virus nucleoprotein, and preparation method thereof
  • Time resolved immunofluorescence detection kit for rabies virus nucleoprotein, and preparation method thereof

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Embodiment Construction

[0032] Rabies virus nucleoprotein time-resolved immunofluorescence detection kit is a double-antibody sandwich immunofluorescence analysis method, using a reaction plate coated with an anti-rabies virus nucleoprotein monoclonal antibody, adding standards and samples to be tested, and the nuclei in the sample The protein is combined with the coated monoclonal antibody to form an antibody-nucleoprotein complex, and then another strain of anti-rabies virus nucleoprotein monoclonal antibody labeled with lanthanide ions and a chelating agent of lanthanide ions, that is, a fluorescence enhancement solution, are added. Formation of antibody-nucleoprotein-antibody-chelator-lanthanide ion complexes. When the lanthanide ions dissociate from the nucleoprotein monoclonal antibody in the fluorescence enhancement solution, they emit strong fluorescence, and the fluorescence intensity is proportional to the nucleoprotein concentration in the sample. The amount of antigenic nucleoprotein in t...

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Abstract

The invention discloses a time resolved immunofluorescence detection kit for rabies virus nucleoprotein, and a preparation method thereof. The content of rabies virus nucleoprotein in a sample is quantificationally determined by using principles of a double-antibody sandwich method and time resolved immunofluorescence. The detection kit comprises a standard, a coating reaction plate, a lanthanide ion marked anti-rabies virus nucleoprotein monoclonal antibody, an analysis buffer, a washing liquid and a fluorescence enhancing liquid, wherein a first anti-rabies virus nucleoprotein monoclonal antibody is adsorbed on the coating reaction plate, and a second anti-rabies virus nucleoprotein monoclonal antibody is marked with the lanthanide ion. A preparation method comprises: adding a to-be measured sample in the coating reaction plate, incubating, washing, adding the lanthanide ion marked second anti-rabies virus nucleoprotein monoclonal antibody, incubating, washing, adding the fluorescence enhancing liquid, and determining in a time resolved immunofluorescence detector. The technical scheme of the invention has advantages of high sensitivity, good singularity, simple operation, wide detection scope and the like.

Description

technical field [0001] The invention relates to the technical field of virus protein immune analysis, in particular to a rabies virus nucleoprotein time-resolved immunofluorescence detection kit and a preparation method thereof. Background technique [0002] Rabies Virus (RV) belongs to Rhabdoviridae, genus Rabies, bullet-shaped, 180nm long, 75nm in diameter, its head is hemispherical, and the end is usually flat. The virus particle consists of two parts: the shell and the core. . The rabies virus genome consists of 11,928 to 11,932 nucleotides. It is a single-stranded, non-segmented, negative-strand RNA that contains 5 large open reading frames and encodes 5 structural proteins. The order of the genome from the 3' end to the 5' end is N, P, M, G and L genes, which encode viral nucleoprotein (NP), phosphoprotein (NS), membrane matrix protein (matrix protein, MP) respectively. ), glycoprotein (glycoprotein, GP) and transcription large protein (polymerase, LP). Studies ...

Claims

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Application Information

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IPC IPC(8): G01N33/577G01N21/64
Inventor 方浩霖刘洁萍何春辉
Owner GUANGZHOU PROMISE BIOLOGICAL PROD
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