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Kit for inspecting infection of swine pleuropneumonia actinomyces

A technology for actinobacillus and pleuropneumonia, which is applied in the field of kits for detection of wild virus infection of actinobacillus pleuropneumoniae, can solve problems such as incompatibility, low vaccine immunity, failure, etc., and achieve good specificity and sensitivity, Convenient collection and purification, good antigenic effect

Inactive Publication Date: 2006-12-27
JIANGSU ACAD OF AGRI SCI
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] App has as many as 12 serotypes, and there is insufficient cross-protection between different serotypes. In addition, the pathogen is prone to drug resistance. Therefore, vaccine immunization is the most effective way to prevent this disease, but in different countries and regions The prevailing serotypes may be different, that is, there may be multiple serotypes in the same pig farm; at present, the whole-bacteria inactivated vaccine containing some of the 12 serotypes is widely used at home and abroad, so there will be some The App multivalent vaccine used in pig farms does not match the actual circulating serotype, resulting in low immunity or failure of the vaccine, causing huge economic losses to pig farms

Method used

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  • Kit for inspecting infection of swine pleuropneumonia actinomyces
  • Kit for inspecting infection of swine pleuropneumonia actinomyces
  • Kit for inspecting infection of swine pleuropneumonia actinomyces

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Embodiment Construction

[0034] The specific embodiments of the present invention will be described in further detail below in conjunction with the accompanying drawings

[0035] 1. Primer design

[0036] According to the ApxIV sequence (sequence number: AF021919) of the App1 type (shope strain purchased from Qingdao Institute of Agriculture, Ministry of Agriculture) published by GenBank and analyzed by the application DNAStar software, the 3'end of the ApxIV gene was selected from 5434bp-6511bp (ApxIV C-terminal antigenicity is more Strong 359 amino acids), design a pair of specific primers by ourselves, add restriction enzyme sites BamHI and HindIII and protective bases at both ends of the primers, and the underlined part is the enzyme site.

[0037] P 1 : 5’-gca ggatcc aaatttaccgatgtg-3’ BamHI

[0038] P 2 : 5’-gta aagctt cctcttcaagcgacaca-3’ HindIII

[0039] 2. Get ApxIV target gene by PCR

[0040] PCR reaction conditions: preheating at 94°C for 5 min, denaturing at 94°C for 1 min, annealing at 60°C ...

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Abstract

A reagent kit consists of enzyme labeled board, IgG antibody, citric acid sodium bicarbonate, colour display system, positive and negative serum. It is featured as using PCR to amplify target gene of ApxIV gene, forming expression plasmid PET32a-ApexIV containing target gene, converting plasmid to be host bacterium BL21(DE3) and purifying in vitro expression protein to be antigen, setting up ELISA detection method and optimizing reaction condition to form the reagent kit.

Description

1. Technical Field [0001] The invention relates to a kit for detecting swine Actinobacillus pleuropneumoniae wild virus infection, which belongs to the development of an animal antigen antibody detection kit, and is dedicated to the detection, epidemiological investigation and immune monitoring of Actinobacillus pleuropneumoniae (App) Wait. 2. Background technology [0002] Porcine infection pleuropneumonia (porcine infection pleuropneumonia) is a highly infectious respiratory disease in pigs caused by Actinobacillus pleuropeumoniae (App), with hemorrhagic, necrotizing pneumonia and chronic fibrinous pleuropneumonia as examples feature. The disease has become one of the three major respiratory infectious diseases in today's large pig farms. Compared with other respiratory diseases, its harmfulness lies in its high morbidity and mortality, especially in newly introduced pig herds. The acute mortality rate can reach 80-100%. [0003] App has many virulence factors, including capsul...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/569C12N15/31C12Q1/04G01N33/543G01N33/535C07K14/36C07H19/00C07H21/00G01N33/52
Inventor 何孔旺彭小华王芳邱索平张雪寒倪艳秀郭容利俞正玉陆承平
Owner JIANGSU ACAD OF AGRI SCI
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