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Reagent box for inspecting infection of swine pleuropneumonia actinomyces

A technology for actinobacillus and pleuropneumonia, which is applied in the field of kits for detection of porcine actinobacillus pleuropneumoniae wild virus infection, 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: 2005-07-27
JIANGSU ACADEMY OF AGRICULTURAL SCIENCES
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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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  • Reagent box for inspecting infection of swine pleuropneumonia actinomyces
  • Reagent box for inspecting infection of swine pleuropneumonia actinomyces
  • Reagent box for inspecting infection of swine pleuropneumonia actinomyces

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Embodiment Construction

[0034] The specific embodiment of the present invention is described in further detail below in conjunction with accompanying drawing

[0035] 1. Primer Design

[0036]According to the ApxIV sequence (serial number: AF021919) of the App1 type published by GenBank (the shope strain was purchased from the Qingdao Animal Inspection Institute of the Ministry of Agriculture), the DNAStar software was used to analyze and select the 3' end 5434bp-6511bp of the ApxIV gene (the antigenicity of the ApxIV C-terminal Strong 359 amino acids), a pair of specific primers were designed by ourselves, restriction enzyme cutting sites BamHI and HindIII and protective bases were added to the two ends of the primers, and the underlined part was the restriction enzyme cutting site.

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

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

[0039] 2. Obtain ApxIV target gene by PCR

[0040] PCR reaction conditions: preheating at 94°C for 5...

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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 wild-virus infection of Actinobacillus pleuropneumoniae, which belongs to the development of a detection kit for animal antigen antibodies, and is specially used for the detection, epidemiological investigation and immune monitoring of Actinobacillus pleuropneumoniae (App). Wait. 2. Background technology [0002] Porcine infectious pleuropneumonia (porcine infection pleuropneumonia) is a highly contagious respiratory disease of pigs caused by Actinobacillus pleuropneumoniae (App), mainly characterized by hemorrhagic, necrotizing pneumonia and chronic fibrinous pleuropneumonia. feature. This disease has become one of the three major respiratory infectious diseases in today's large pig farms. Compared with other respiratory diseases, its harmfulness is that it has high morbidity and mortality, especially in newly introduced pigs. The acute mortality rate can reach 80-100%. [0003] App has many virulence...

Claims

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Application Information

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IPC IPC(8): C07H19/00C07H21/00G01N33/52G01N33/535G01N33/543G01N33/569
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 ACADEMY OF AGRICULTURAL SCIENCES
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