Salmonella cholerae enzyme-linked immunosorbent assay kit

An enzyme-linked immunosorbent reagent, Salmonella technology, applied in the field of biotechnology and immunology, can solve the problem of weak resistance to disinfectants

Inactive Publication Date: 2017-04-26
JIANGSU WISE SCI & TECH DEV
View PDF2 Cites 2 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

It can survive for several weeks, months or even years in suitable organic matter, but

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
  • Salmonella cholerae enzyme-linked immunosorbent assay kit
  • Salmonella cholerae enzyme-linked immunosorbent assay kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0056] Example 1. Anti-Salmonella choleraesuis monoclonal antibody 5D6D9G3B4 and 5D2G2D9C1 preparation of

[0057] 1. Preparation of immunogen and positive standard

[0058] Salmonella choleraesuis (ATCC No.27660) was inoculated in Brain Heart Infusion Broth (BHI), cultured with shaking at 150r / min for 17h, counted, and inactivated by adding 0.3% formaldehyde solution at room temperature for 1 day. Adjust the concentration of Salmonella choleraesuis (ATCC No. 27660) to 5×10 with normal saline 9 cfu / ml as the immunogen; adjust the concentration to 10 with saline 8 Cfu / ml was used as a positive control standard, and brain heart extract broth (BH workers) was used as a negative control standard.

[0059] 2. Preparation of monoclonal antibodies

[0060] 1) Experimental animals: Three 8-week-old, weighing about 20 g female Balb / c mice were selected as experimental animals.

[0061] 2) Immunization method: each mouse was intraperitoneally injected with 0.2 ml of imm...

Embodiment 2

[0086] Example 2. Characterization of monoclonal antibodies 5D6D9G3B4 and 5D2G2D9C1

[0087] 1. Monoclonal Antibody Subclass Identification

[0088] 1. Antigen coating: Coat goat anti-mouse secondary antibody IgG+A+M with 0.01MPBS, 50 μL per well, coat overnight at 4°C, discard the liquid in the well the next day, and wash the plate 3 times.

[0089] 2. Blocking: add 200 μL of 1% BSA to each well, and block overnight at 4°C. Pat the board dry the next day without washing it.

[0090] 3. Add monoclonal antibody hybridoma cell supernatant, each sample has 8 microwells, 50 μL per well, and incubate at 37°C for 1 hour.

[0091] 4. After washing the plate 4 times, add specific binding rabbit anti-mouse IgG1, IgG2a, IgG2b, IgG3, IgA, IgM, and incubate at 37°C for 1 hour.

[0092] 5. After washing the plate 4 times, add diluted horseradish peroxidase-labeled anti-rabbit secondary antibody IgG (H+L) to each well, and incubate at 37°C for 30 minutes.

[0093] 6. After washing the p...

Embodiment 3

[0102] Example 3. The composition, preparation and application of the ELISA kit for detecting Salmonella choleraesuis

[0103] 1. The enzyme-linked immunosorbent assay kit consists of the following materials:

[0104] (1) Antibody pre-coated microtiter plate: Dilute with 0.02M acetate buffer (pH2.0) solution, and coat 96-well microtiter plate with anti-Salmonella choleraesuis monoclonal antibody 5D6D9G3B4, 100miu5L per well. Incubate overnight at 4°C, block and wash according to conventional ELisa methods.

[0105] (2) Salmonella choleraesuis positive control standard and negative control standard.

[0106] (3) Anti-Salmonella choleraesuis monoclonal antibody 5D2G2D9C1 labeled with horseradish peroxidase.

[0107] (4) Enzyme-labeled antibody diluent: 0.01M PBS, pH7.6.

[0108] (5) 10× concentrated lotion: 0.1M phosphate buffer containing 0.5% Tween 20 and 0.2% sodium azide, pH 7.4, just dilute the concentrated lotion 10 times before use.

[0109] (6) Color developing solut...

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 an enzyme-linked immunosorbent assay kit for detecting salmonella cholerae. The kit comprises two monoclonal antibodies which can be specifically combined with salmonella cholerae, wherein one is a monoclonal antibody 5D6D9G3B4 exclusively serving as a capture antibody, and the other one is a monoclonal antibody 5D2G2D9C1 serving as a detection antibody. A large number of tests prove that the kit can specifically and efficiently detect salmonella cholerae and does not carry out cross reaction with other 74 common pathogenic bacteria, and is a pathogenic bacteria detection product with perfect performance.

Description

technical field [0001] The invention belongs to the fields of biotechnology and immunology, and in particular relates to an enzyme-linked immunoassay kit for detecting Salmonella choleraesuis. Background technique [0002] There are more than 2,000 serotypes of Salmonella, and more than 200 serotypes have been found in China. Salmonella is a Gram-negative bacillus that can move, does not form spores, is facultatively anaerobic, and has flagella all over the body. The optimum growth temperature is 37°C, and the optimum pH is 6.8-7.4. Salmonella has tenacious vitality, strong resistance to the outside world, and has certain resistance to factors such as drying and corruption. It can reproduce at 7-45°C, and can be frozen or It survives after freeze-drying, can survive the winter in frozen soil, can survive in pig manure for 1-8 months, can survive in manure oxidation pond for 47 days, can survive in bedding grass for 2-5 months, and can survive in 10% -19% can survive more t...

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): G01N33/577G01N33/558
CPCG01N33/558G01N33/56916G01N33/577
Inventor 杜霞刘静张淑雅
Owner JIANGSU WISE SCI & TECH DEV
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