Envision immunohistochemical detection method of Clostridum Perfringens alpha toxin

A clostridium perfringens and immunohistochemical technology, applied in the field of animal bacteriology, can solve the problems of inaccurate results, time-consuming and laborious, and achieve the effects of good repeatability, low background, and long-term preservation.

Inactive Publication Date: 2016-11-09
SHANDONG AGRICULTURAL UNIVERSITY
View PDF1 Cites 5 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Clostridium perfringens is mainly an enterogenous infection, which often forms enterogenous toxemia, but does not necessarily form bacteremia and sepsis. Therefore, bacteria may not be isolated from visceral parenchymal organs; When experimental animals do toxin inoculation experiments and toxin neutralization experiments, standard positive hyperimmune serum for toxins is required, which is time-consuming and laborious, and due to individual differences in animals, it can also lead to inaccurate results

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
  • Envision immunohistochemical detection method of Clostridum Perfringens alpha toxin
  • Envision immunohistochemical detection method of Clostridum Perfringens alpha toxin

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0064] Preparation of Clostridium perfringens alpha toxin:

[0065] Type A Clostridium perfringens strains (National Type Culture Collection, British National Collection Center Preservation Number: NCTC528) were inoculated on the blood plate medium for resuscitation, anaerobic culture at 37°C for 36 hours, and single colonies were picked and added to thioglycolic acid Salt enrichment culture medium, at a gas concentration of 88% N 2 , 7%H 2 , 5%CO 2 Incubate at 38°C for 12 hours under anaerobic conditions. Inoculate 5mL type A Clostridium perfringens enrichment solution into 100ml pH7.5 toxin-producing medium (dissolve 3g peptone, 1.2g dextrin, 2g yeast extract and 2.2gL-arginine per 100mL PBS buffer acid), shake culture in an anaerobic environment, culture at 43°C for 5h to efficiently produce toxin, then centrifuge at 3800g at 4°C for 15min, and then filter and sterilize with a Chua filter with a pore size of 0.22μm to obtain α-toxin as The exotoxin of the Lord. Using l...

Embodiment 2

[0067] Artificial Infection of Clostridium perfringens in Rabbits

[0068] Select 6 rabbits of 1.0-1.5Kg just after weaning, and randomly divide them into experimental group and control group, 3 / group, and the experimental group intramuscularly injects the above-mentioned bacteria-eliminated mice to measure the LD50 of 2 4.25 3 mL of α-toxin, and the control group was injected with the same amount of normal saline, and the disease state of the animals was observed within 72 hours. At the time of death, the lesion was immediately clinically dissected, and the heart, liver, spleen, lung, kidney, stomach, small intestine and cecum were taken from the diseased tissue, and put into the improved Bouin'S liquid fixative for subsequent preparation of tissue slices for positive control and Immunohistochemical staining, the negative control group was carried out at the same time;

[0069] The composition of the improved Bouin'S liquid fixative described therein is: 750mL saturated picr...

Embodiment 3

[0071] Preparation of tissue sections

[0072] After the tissue was fixed in solution for 48h, the tissue was cut into 1cm with a blade 3 When the tissue block was washed with running water until there was no yellow fixative solution, it was washed with 70% alcohol (overnight), 80% alcohol (1h), 85% alcohol (1h), 90% alcohol (1h), 95% alcohol ( 1h), 100% alcohol (45min), xylene: alcohol (10min) gradient dehydration, xylene transparent tissue 10s, 60 ℃ immersion in wax for 1.5h, paraffin embedding, sliced ​​into 4um thick slices by a microtome, sliced ​​at 37 Dry in an incubator at ℃, place at 4℃ for subsequent immunohistochemical staining, and perform this operation on the negative control group at the same time.

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 relates to the field of animal bacteriology, and provides an Envision immunohistochemical detection method of Clostridum Perfringens alpha toxin. The Envision immunohistochemical detection with the advantages of high specificity, good repeatability, low background and realization of visual and simple detection of the alpha toxin is established on the basis of a mouse anti-alpha toxin monoclonal antibody. The method can be used for detecting the Clostridum Perfringens alpha toxin in tissues of animals except mice, and provides an effective and reliable way for detection of the distribution of the toxin in the animal bodies.

Description

technical field [0001] The invention relates to the field of animal bacteriology and provides an immunohistochemical detection method of Clostridium perfringens alpha toxin Envision. Background technique [0002] Clostridium perfringens can produce a variety of exotoxins. According to the four main lethal exotoxins it produces, Clostridium perfringens can be divided into A(α), B(α, β, ε), C(α , β), D (α, ε) and E (α, ι) types, each type produces α toxin, which has two kinds of toxicity, lecithinase C and sphingomyelinase, and can simultaneously hydrolyze phosphatidyl bile on the cell membrane Alkali and sphingomyelin components, destroy the cell membrane structure, lead to cell lysis, thus presenting cytotoxicity, hemolysis, lethality and skin necrosis, etc., in animal necrotic enteritis, enterotoxemia, traumatic gas gangrene of humans and animals Play a key role, causing huge losses to the breeding industry and public health. [0003] At present, the detection methods of ...

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): G01N33/577
CPCG01N33/543G01N2333/33
Inventor 王海荣秦贵平柴同杰陈长俊
Owner SHANDONG AGRICULTURAL UNIVERSITY
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