Method for detecting bovine brucellosis by competence ELISA (Enzyme-Linked Immunosorbent Assay) kit

A technology of Brucella bovis and Brucella bovis, applied in the field of biotechnology detection, can solve problems such as missed detection and wrong detection

Active Publication Date: 2014-08-20
YANGZHOU UNIV
View PDF4 Cites 1 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] So far, monoclonal antibodies against the A antigen, M antigen, C antigen and C/Y antigen located on the lipopolysaccharide O chain (O-LPS) have been prepared; on the other hand, scholars have found that some specific outer membrane proteins There is also immunogenicity, such as OMP31 and other cELISA methods establ...

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 detecting bovine brucellosis by competence ELISA (Enzyme-Linked Immunosorbent Assay) kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment Construction

[0022] 1. Monoclonal antibody bru-5C10 with good reactivity to Brucella bovis was obtained by cell fusion technology:

[0023] The splenocytes and SP2 / 0 myeloma cells of mice after routine immunization were used for cell fusion according to standard procedures. The supernatant of hybridoma cells was collected and screened by indirect enzyme-linked immunosorbent assay (iELISA) to have a strong reaction with LPS crudely extracted from Brucella bovis A99, but not with LPS crudely extracted from Yersinia enterocolitica O9. Reactive positive hybridoma cell lines. The monoclonal antibody bru-5C10 was obtained by subcloning twice according to the limiting dilution method.

[0024] 2. Preservation certificate information of monoclonal antibody bru-5C10:

[0025] The deposit number is CGMCC No.9219, and the deposit address is: No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing, China General Microorganism Culture Collection Management Center, deposit date: May 7, 2014. Bio...

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 detecting bovine brucellosis by a competence ELISA (Enzyme-Linked Immunosorbent Assay) kit, belonging to the field of biotechnical detection. The method comprises the steps: coating bovine brucella A99 subjected to ultrasonic splitting by an ELISA enzyme-labeled plate; then adding an anti-brucella specific monoclonal antibody bru-5C10 and a diluted to-be-detected serum sample in plate holes of the ELISA coated plate; and simultaneously respectively adding the anti-brucella specific monoclonal antibody bru-5C10 and known negative and positive serum in the plate holes of the ELISA enzyme-labeled plate, washing with PBST (polybutylene terephthalate) after water bathing, respectively adding an anti-mouse IgG (Intravenous Gamma Globulin) enzyme-labeled antibody in each plate hole, performing a chromogenic reaction after incubation and washing, measuring an OD450 value of each plate hole, and calculating an inhibition rate of each plate. According to the method, yersinia enterocolitica O9 and escherichia coli O157 high-immunity positive serum are detected by using an established cELISA method, a PI (inhibition rate) value is lower than 30 percent, and the generation of a false positive result can be effectively eliminated.

Description

technical field [0001] The invention belongs to the field of biotechnology detection, in particular to the technical field of detection of brucellosis in cattle and sheep. Background technique [0002] Brucellosis (brucellosis) is an important zoonotic disease caused by Brucella, and ruminants, especially cattle and sheep, are susceptible to infection. According to the antigen type and host type, Brucella can be divided into 6 types. The general symptom of sick female animals is abortion, while males can cause reproductive disorders such as infertility. In recent years, brucellosis has shown a rebound trend worldwide. In China, the damage caused by B. abortus and B. melitensis is particularly extensive and serious. The disease has caused huge economic losses to animal husbandry and public health, and it is of great practical significance to strengthen the prevention and control of brucellosis. [0003] Accurate diagnosis of brucellosis is the primary measure to prevent the...

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
IPC IPC(8): G01N33/577
CPCG01N33/54373G01N33/56911G01N2333/23
Inventor 秦爱建王志明丁家波邵红霞钱琨冯忠武
Owner YANGZHOU UNIV
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