Method for united typing detection of porcine contagious pleuropneumonia antibody and kit

A combined detection technology for pleuropneumonia, applied in measurement devices, analytical materials, material stimulation analysis, etc., can solve the problems of high cost, large amount of serum used, time-consuming App typing detection, etc., and achieve good stability and specificity Strong, specificity-enhancing effect

Inactive Publication Date: 2010-10-13
SHANGHAI ENTRY EXIT INSPECTION & QUARANTINE BUREAU OF P R C
View PDF6 Cites 2 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The technical problem to be solved by the present invention is to overcome the deficiencies in the prior art, and provide a method for simultaneous typing and detection of porcine infectious pleuropneumonia antibodies. The establishment of this method can meet the requirements of import and export breeding pig quarantine, fast and accurate, and at the same time, provide a new applicable method for the prevention and control of porcine infectious pleuropneumonia in my country's pig industry

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Method for united typing detection of porcine contagious pleuropneumonia antibody and kit
  • Method for united typing detection of porcine contagious pleuropneumonia antibody and kit
  • Method for united typing detection of porcine contagious pleuropneumonia antibody and kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0038] 1. Preparation of S1-S7 polysaccharide antigens of Actinobacillus pleuropneumoniae

[0039] The App serum S1-S7 type standard reference strains (gifted by the Danish National Veterinary Research Institute) which were lyophilized and stored were streaked and inoculated in the 0.1% nicotine adenine dinucleotide (NAD, purchased from Sinopharm Chemical Reagent Co., Ltd. ) TSCA medium (the TSCA medium is prepared by our laboratory: take 40g TSA (Tryptic Soy Agar, DIFCO TM ) agar powder, dissolved in 1000mL deionized water, stirred and heated to dissolve, boiled for 1min until completely dissolved, then autoclaved at 121°C for 15min, cooled to 40°C-50°C, added 8% (V / V) defibrillated sheep blood ( purchased from Zhuzhai Sterile Animal Blood Reagent Supply Station, Minhang District, Shanghai) and 0.15% glucose (W / V), in a water bath at 80°C to 82°C for 10min to 12min, the color of the culture medium changed from blood red to chocolate, and appeared fine particles. Cool to abo...

Embodiment 2

[0055] Example 2 Liquid phase protein chip specific detection

[0056] Use liquid chip typing to detect porcine infectious pleuropneumonia antibody test method to detect App serum type 1-7 pig positive sera, porcine PRRS positive sera (both European strains and American strains come from the National Veterinary Laboratory of the United States); pig rings Virus positive serum, Haemophilus parasuis positive serum, Toxoplasma porcine positive serum, Mycoplasma pneumoniae positive serum (gifted by Shanghai Animal Disease Prevention and Control Center); Porcine O-type foot-and-mouth disease positive serum (purchased from Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences) ); porcine transmissible gastroenteritis positive serum (svanova biotech ELISA kit); porcine brucellosis positive serum (purchased from China Veterinary Drug Control Institute); porcine Streptococcus suis positive serum (gifted by Shanghai Jiaotong University Agricultural College); porc...

Embodiment 3

[0059] Example 3 Liquid-phase chip detection method and ELISA detection sensitivity comparison

[0060] The App S1-S7 type positive pig serum was diluted 1:200-1:25600, and the liquid chip was used to detect the antibody of porcine infectious pleuropneumonia. The detected titer is generally 2-4 dilutions higher than that of ELISA, see Table 3 for details.

[0061] Table 3 Comparison between liquid phase chip (referred to as liquid core) and ELISA detection kit

[0062]

[0063]

[0064] Note: +: positive; -: negative; ±: suspicious

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

PUM

No PUM Login to view more

Abstract

The invention discloses a method for united typing detection of a porcine contagious pleuropneumonia antibody. The method comprises the following steps: firstly preparing actinobacillus pleuropneumoniae polysaccharide antigen and rabbit anti-actinobacillus pleuropneumonia hyper-immune serum, and then purifying; coupling a liquid-phase chip microballoon by utilizing the purified rabbit anti-actinobacillus pleuropneumoniae hyper-immune serum, building a method for typing detection of the liquid-phase chip of the porcine pleuropneumonia antibody according to a double-sandwich ELISA principle, and determining the optimum experimental condition; and finally determining the threshold for positive and negative judgment of the liquid-phase chip through statistical analysis. In addition, the invention also discloses a kit for united typing detection of the liquid-phase chip of the porcine contagious pleuropneumoniae antibody. The invention can simultaneously carry out typing detection of the S1-S7-type serum antibody of porcine contagious pleuropneumonia, and the whole reaction can be completed within 3 hours; and the method has the characteristics of rapidly, sensitively, specifically andsimultaneously detecting a plurality of serum types, thus the method can be used for preliminarily screening entry and exit boars and diagnosing and monitoring porcine contagious pleuropneumoniae in hogpens of China.

Description

technical field [0001] The invention relates to the technical fields of agriculture and animal quarantine, in particular to a method for combined typing and detection of porcine infectious pleuropneumonia antibodies; in addition, the invention also relates to a liquid-phase chip combined typing detection reagent for porcine infectious pleuropneumonia antibodies box. Background technique [0002] Porcine contagious pleuropneumoniae (Porcine contagious pleuropneumoniae) is a respiratory infectious disease of pigs caused by Actinobacillus pleuropneumoniae (App for short). Its morbidity and mortality are all above 50%, and the mortality rate of the most acute type can be as high as 80-100%. The chronic type can cause pigs to grow slowly and become a dead pig. This disease is one of common pig diseases in intensive pig farms. The disease is one of the swine diseases that must be checked for imported breeding pigs in many countries. my country has repeatedly detected porcine infe...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

Application Information

Patent Timeline
no application Login to view more
Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/569G01N33/543G01N21/64
Inventor 李树清王艳王巧全张强杜军陈志飞
Owner SHANGHAI ENTRY EXIT INSPECTION & QUARANTINE BUREAU OF P R C
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap