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Antigen for detecting porcine infectious pleuropneumonia antibody and preparation method of antigen

A porcine pleuropneumonia and pleuropneumonia technology is applied in the field of antigens and preparations for porcine infectious pleuropneumonia antibody detection, which can solve the problems of inability to meet clinical testing requirements, lack of cross-protection, cumbersome operation, etc., and achieve good detection effect, strong specificity, Simple operation effect

Active Publication Date: 2015-04-08
YEBIO BIOENG OF QINGDAO
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, due to the lack of cross-protection between APP types, if each sample is tested for 15 serotypes, the operation is cumbersome, the workload is heavy, it is difficult to implement, and it cannot meet the needs of clinical testing.

Method used

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Examples

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preparation example Construction

[0014] The preparation method of the antigen of the present invention is as follows:

[0015] 1 Preparation of seeds for antigen production

[0016] 1.1 Preparation of chicken broth agar (referred to as S agar) Take 100ml of chicken broth, 3g of tryptone blood agar matrix, 0.5g of polypeptone, heat and dissolve, adjust the pH value to 7.2-7.4, sterilize at 115°C for 30 minutes, and cool to 55°C At about ℃, add 5ml of inactivated chicken serum and 1ml of 1% coenzyme I (NADH) according to the sterile requirements.

[0017] 1.2 Primary seed propagation and identification Streak inoculation of APP1-15 freeze-dried strains on S agar plate, place in 5%-10% CO 2 Under the conditions, culture at 37°C for 16-18 hours, select more than 5 typical colonies (or bacterial lawns) with iridescent luster, inoculate them into the yolk sac of SPF chicken embryos aged 5-6 days, continue to incubate at 37°C, and collect them within 30 hours The yolk fluid of dead chicken embryos shall be regarde...

Embodiment 1

[0037] 1 Preparation of seeds for antigen production

[0038] 1.1 First-level seed propagation and identification Streak inoculation of APP1-15 freeze-dried strains on S agar plate, place in 5%-10% CO 2 Under the conditions, culture at 37°C for 16-18 hours, select more than 5 typical colonies (or bacterial lawns) with iridescent luster, inoculate them into the yolk sac of SPF chicken embryos aged 5-6 days, continue to incubate at 37°C, and collect them within 30 hours The yolk fluid of dead chicken embryos shall be regarded as first-class seeds after passing the pure inspection. Stored at -20°C, the usable period should not exceed 1 month; stored below -70°C, the usable period should not exceed 3 months, and the subculture on the medium should not exceed 6 generations.

[0039] 1.2 Propagation of secondary seeds Take primary seeds of various types of strains, inoculate S agar plate by streaking, and inoculate with 5% to 10% CO 2 Under the conditions, cultivate at 37°C for 16...

Embodiment 2

[0051] 1 Preparation of seeds for antigen production

[0052] 1.1 First-level seed propagation and identification Streak inoculation of APP1-15 freeze-dried strains on S agar plate, place in 5%-10% CO 2 Under the conditions, culture at 37°C for 16-18 hours, select more than 5 typical colonies (or bacterial lawns) with iridescent luster, inoculate them into the yolk sac of SPF chicken embryos aged 5-6 days, continue to incubate at 37°C, and collect them within 30 hours The yolk fluid of dead chicken embryos shall be regarded as first-class seeds after passing the pure inspection. Stored at -20°C, the usable period should not exceed 1 month; stored below -70°C, the usable period should not exceed 3 months, and the subculture on the medium should not exceed 6 generations.

[0053] 1.2 Propagation of secondary seeds Take primary seeds of various types of strains, inoculate S agar plate by streaking, and inoculate with 5% to 10% CO 2 Under the conditions, cultivate at 37°C for 16...

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Abstract

The invention provides an antigen for detecting a porcine infectious pleuropneumonia antibody of swinery and a preparation method of the antigen. The antigen is a polysaccharide antigen which is extracted from type 1-15 strain cultures of porcine infectious pleuropneumonia actinobacillus; the polysaccharide antigen is a bacterial excreted polysaccharide which has very strong specificity and sensibility and is capable of truly detecting the porcine infectious pleuropneumonia antibody; the determination of a normal concentration critical value of the antigen provides guarantee for the detection of enzyme linked immunosorbent assay and the detection of indirect hemagglutination test; the preparation method of the antigen is simple to operate, convenient and quick, and suitable for popularization and application at the basic level, and does not need any special equipment and instrument.

Description

technical field [0001] The invention belongs to the technical field of veterinary biological products, and in particular relates to an antigen for detecting porcine infectious pleuropneumonia antibody and a preparation method thereof. Background technique [0002] Porcine infectious pleuropneumonia is an important respiratory infectious disease of pigs caused by Actinobacillus pleuropneumoniae (APP). This disease is mainly caused by a fibrinous, hemorrhagic, and necrotizing pneumonia accompanied by pleurisy in pigs. Most of them are the most acute or acute course of the disease and they die quickly. The morbidity and mortality are more than 50%. It can occur in any In pigs of the age, porcine infectious pleuropneumonia has occurred in countries all over the world, and the positive rate is increasing year by year. In recent years, there have been reports of this disease occurring in almost all countries and provinces in the country. Especially in intensive pig farms, once it...

Claims

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Application Information

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IPC IPC(8): G01N33/531G01N21/33
CPCG01N33/531G01N33/56911G01N2333/195
Inventor 马爽范根成孙健宋新宇郭玉广郭莉莉胡潇程增青王红李金积
Owner YEBIO BIOENG OF QINGDAO
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