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Chemiluminescence immunoassay kit for Seneca valley virus nonstructural protein 3ABC antibody detection

A chemiluminescence immunoassay, non-structural protein technology, applied in chemiluminescence/bioluminescence, analysis by chemical reaction of materials, material analysis by optical means, etc., to achieve high protein yield, simple operation, and improved detection sensitivity. Effect

Inactive Publication Date: 2018-11-23
LANZHOU INST OF VETERINARY SCI CHINESE ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

In this new discovery that describes how certain viruses cause diseases called Sickness Fever (SF) have been discovered by researchers at home or abroad. These symptoms are caused when they infect humans through contact between people who come into direct physical proximate during their interactions over long periods without being sickened from them. They also affect other animals like dogs but it's important because these harmful agents may spread quickly throughout human populations worldwide. Current methods require expensive equipment such as ELISA analysis, making testing difficult due to limited resources available.

Problems solved by technology

The problem addressed in this patented text relates to identifying and differentiate Swine Senkevices Virus Variant Causing Foot Blotch Syndrome (SFRS); specifically, finding ways to identify and separate these viruses without causing damage to humans.

Method used

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  • Chemiluminescence immunoassay kit for Seneca valley virus nonstructural protein 3ABC antibody detection
  • Chemiluminescence immunoassay kit for Seneca valley virus nonstructural protein 3ABC antibody detection
  • Chemiluminescence immunoassay kit for Seneca valley virus nonstructural protein 3ABC antibody detection

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Effect test

Embodiment 1

[0049] Example 1 Preparation and assembly of chemiluminescent immunoassay kit for detection of Seneca Valley virus nonstructural protein 3ABC antibody

[0050] (1) Preparation of the kit:

[0051] 1. Expression of Seneca Valley virus nonstructural protein 3ABC antigen:

[0052] The 969bp gene encoding the Seneca Valley virus nonstructural protein 3ABC (shown in SEQ ID NO.1) was cloned into the pET-30a(+) prokaryotic expression vector. After digestion and sequencing were correct, the positive plasmid was transformed into the BL21DE3plysS strain, Kanamycin-resistant LB plates were used to screen single clones, shake the LB medium at 37°C overnight; transfer the overnight bacterial solution to fresh LB medium at a volume ratio (v / v) of 1:100, shake at 37°C at 200rpm After 3 hours until the OD600 value reaches about 0.6, add IPTG with a final concentration of 1 mmol / L to continue shaking the bacteria for 3 hours, and collect the bacterial liquid by centrifugation at 6000 rpm. Re...

Embodiment 2

[0089] Example 2 Sensitivity and specificity experiments of the chemiluminescent immunoassay kit for detection of Seneca Valley virus nonstructural protein 3ABC antibody

[0090] 1. Dilution of known background serum:

[0091] (1) A total of 30 serum samples were obtained from clinically healthy pigs that had not been infected with Seneca Valley virus. These sera were used to evaluate the specificity of the chemiluminescent detection method for antibodies to nonstructural proteins of Seneca Valley virus.

[0092] (2) 30 samples each of porcine vesicular disease positive serum, classical swine fever virus positive serum, and porcine circovirus-2 positive serum, these sera were used to evaluate the specificity of the chemiluminescent detection method for Seneca Valley virus nonstructural protein 3ABC antibody .

[0093] (3) A total of 80 serum samples were from pigs challenged experimentally with SVV. The pigs had not received any immunizations prior to the challenge and these...

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Abstract

The invention discloses a chemiluminescence immunoassay kit for Seneca valley virus nonstructural protein 3ABC antibody detection. The kit comprises a Seneca valley virus nonstructural protein 3ABC antigen coated chemiluminescence immunoassay plate, positive control serum, negative control serum, an enzyme-labeled antibody, a sample diluent, a chemiluminescence substrate solution, a luminescence enhancing agent and concentrated washing liquid, wherein the amino acid sequence of the Seneca valley virus nonstructural protein 3ABC is shown as SEQ ID NO.2. The kit provided by the invention uses nonstructural protein 3ABC antigen expressed by a prokaryotic expression system for coating a reaction plate; the antigen consumption is low; whether the Seneca valley virus exists in the pig serum or not can be efficiently detected; in addition, reaction with the swine vesicular disease viruses, hog cholera viruses and porcine circovirus type 2 cannot occur. An enzymatic chemiluminescence reactionsystem is used for judging the result; the detection sensitivity is improved. The kit provided by the invention has the advantages that the specificity is high; sensitivity and high efficiency are realized; good market prospects are realized.

Description

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Claims

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Application Information

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Owner LANZHOU INST OF VETERINARY SCI CHINESE ACAD OF AGRI SCI
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