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Enzyme linked immunosorbent assay (ELISA) kit for detecting encephalomyocarditis virus (EMCV) antibodies and application of kit

A technology of enzyme-linked immunosorbent reagents and kits, which is used in measurement devices, instruments, scientific instruments, etc., can solve the problems of low sensitivity and specificity, not widely used, and low accuracy, and achieves strong specificity and detection cost. Low, high sensitivity effect

Inactive Publication Date: 2014-01-15
NORTHWEST UNIVERSITY FOR NATIONALITIES
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] (1) Too specific and not widely used: existing kits can only detect a single species of animal or human;
[0007] (2) The sensitivity and specificity are not high;
[0008] (3) There are many false positives and the accuracy is not high

Method used

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  • Enzyme linked immunosorbent assay (ELISA) kit for detecting encephalomyocarditis virus (EMCV) antibodies and application of kit
  • Enzyme linked immunosorbent assay (ELISA) kit for detecting encephalomyocarditis virus (EMCV) antibodies and application of kit
  • Enzyme linked immunosorbent assay (ELISA) kit for detecting encephalomyocarditis virus (EMCV) antibodies and application of kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0054] 1. Prepare an enzyme-linked immunosorbent assay kit for detecting EMCV antibodies:

[0055] (1) ELISA plate coated with EMCV recombinant antigen;

[0056] (2) Enzyme-labeled EMCV antigen working solution;

[0057] (3) EMCV antibody positive control;

[0058] (4) EMCV antibody negative control;

[0059] (5) Lotion;

[0060] (6) Chromogenic agent A;

[0061] (7) Chromogen B;

[0062] (8) Stop solution.

[0063] (1) Preparation of ELISA plate coated with EMCV recombinant antigen:

[0064] (1) Preparation of EMCV recombinant antigen: mix EMCV-VP1 recombinant protein, EMCV-VP2 recombinant protein, and EMCV-2C recombinant protein at the same protein concentration;

[0065] (2) Coating EMCV recombinant antigen: Dilute the EMCV recombinant antigen protein concentration to 30 μg / ml with pH 9.6, 0.05M sodium carbonate buffer, add 100 μL / well into the microwell of the microplate, place for adsorption, shake Dry;

[0066] (3) Seal the coated EMCV recombinant antigen: u...

Embodiment 2

[0121] The difference between this embodiment and embodiment 1 is:

[0122] 1. Preparation of kits

[0123] (1) Preparation of ELISA plate coated with EMCV recombinant antigen:

[0124] (1) Preparation of EMCV recombinant antigen: mix EMCV-VP1 recombinant protein, EMCV-VP2 recombinant protein, and EMCV-2C recombinant protein at the same protein concentration;

[0125] (2) Coating EMCV recombinant antigen: Dilute the EMCV recombinant antigen protein concentration to 60 μg / ml with pH 9.6, 0.05M potassium carbonate buffer, add 100 μL / well into the microwell of the microplate, place for adsorption, shake Dry;

[0126] (3) Block the coated EMCV recombinant antigen: use 150 μL / well of sodium phosphate buffer solution containing 100 g / L horse serum, 100 g / L sucrose, and pH value of 7.8, and spin dry;

[0127] (4) Drying: After drying at 25°C, put it in a bag containing a desiccant and seal it for storage at 2°C.

[0128] (2) Preparation of enzyme-labeled EMCV antigen workin...

Embodiment 3

[0153] The difference between this embodiment and embodiment 1 is:

[0154] 1. Preparation of kits

[0155] (1) Preparation of ELISA plate coated with EMCV recombinant antigen:

[0156] (1) Preparation of EMCV recombinant antigen: mix EMCV-VP1 recombinant protein, EMCV-VP2 recombinant protein, and EMCV-2C recombinant protein at the same protein concentration;

[0157] (2) Coating EMCV recombinant antigen: Dilute the EMCV recombinant antigen protein concentration to 300ng / ml with pH 9.6, 0.05M sodium carbonate buffer, add 100μL / well into the microwell of the microplate, place for adsorption, shake Dry;

[0158] (3) Seal the coated EMCV recombinant antigen: use sodium phosphate buffer solution containing 5g / L fish peptone, 100g / L sucrose, pH value 7.5, seal at 150μL / well, and spin dry;

[0159] (4) Drying: After drying at 32°C, put it in a bag containing a desiccant and seal it for storage at 8°C.

[0160] (2) Preparation of enzyme-labeled EMCV antigen working solution...

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PUM

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Abstract

The invention discloses an enzyme linked immunosorbent assay (ELISA) kit for detecting encephalomyocarditis virus (EMCV) antibodies and application of the kit. The kit comprises an ELISA plate coating EMCV recombinant antigens, enzyme labeled EMCV antigen working solution, EMCV antibody positive control, EMCV antibody negative control, lotion, a color-developing agent A, a color-developing agent B and stop solution. The invention also relates to application of the kit to detecting EMCV antibodies of human and animals. The kit can detect human and animals, has high sensitivity, strong specificity, good thermal stability, low detection cost and short detection time and is suitable for detecting mass EMCV antibodies. Through tests, positivity and negativity of 99.6% of serums can be directly judged, so the kit has good practical effect.

Description

technical field [0001] The present invention relates to an ELISA kit, in particular to an ELISA kit capable of detecting encephalomyocarditis virus antibodies, and the present invention also relates to the use of the ELISA kit in detecting human and animal encephalomyocarditis virus antibodies application. Background technique [0002] Encephalomyocarditis virus (EMCV) is an important zoonotic pathogen, which can cause pigs and certain mammals, rodents and even primates, and is characterized by encephalitis, myocarditis or pericarditis. acute infectious disease. EMCV infection can cause lethal myocarditis in piglets, causing acute death, with a mortality rate up to 100%, and can lead to abortion, stillbirth, weak fetus, and mummified fetus in pregnant sows; In addition to infecting animals, it has also been detected in humans; in addition, EMCV also exists in the form of subclinical infection. Therefore, it is extremely important to make a good diagnosis and prevention of...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/569
Inventor 冯若飞樊得英马忠仁李明生乔自林李向茸
Owner NORTHWEST UNIVERSITY FOR NATIONALITIES
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