Rabbit Klebsiella pneumoniae agglutinogen and application of rabbit Klebsiella pneumoniae agglutinogen

A Klebsiella and agglutinogen technology, applied in the field of biology, can solve the problems of no research report on Klebsiella pneumoniae microagglutination kit, high technical requirements for operators, and high technical requirements for operators , to achieve the effect of easy observation, high technical requirements, and low technical requirements

Inactive Publication Date: 2012-12-12
CHONGQING ACAD OF ANIMAL SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although the capsular swelling test is fast and convenient, it needs to be observed with the help of a microscope, which cannot be performed in many grassroots veterinary stations and rabbit farms
Although countercurrent immunoelectrophoresis and indirect immunofluorescence tests are accurate, they require higher technical requirements for operators and require more expensive equipment
PCR detection method has high sensitivity and accuracy, but requires expensive instruments and reagents, high detection cost, high technical requirements for operators, and is not suitable for grassroots use
[0004] Through literature search, we have not found relevant research reports on rabbit Klebsiella pneumoniae microagglutination kit and detection method, especially the method of microagglutination test after staining rabbit Klebsiella pneumoniae with fuchsin

Method used

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  • Rabbit Klebsiella pneumoniae agglutinogen and application of rabbit Klebsiella pneumoniae agglutinogen
  • Rabbit Klebsiella pneumoniae agglutinogen and application of rabbit Klebsiella pneumoniae agglutinogen
  • Rabbit Klebsiella pneumoniae agglutinogen and application of rabbit Klebsiella pneumoniae agglutinogen

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] Example 1 Preparation of reagents

[0033] Preparation of a sample serum

[0034] Preparation of serum: the isolated rabbit blood, coagulate naturally, separate the serum, put it in a water bath at 56°C for 30 minutes, and set aside.

[0035] Preparation of diagglutinogen

[0036] Inoculate rabbit Klebsiella pneumoniae on TSA agar plates, place them in a 37°C incubator for 24 hours, wash the lawn with physiological saline; purely test; count live bacteria; adjust the concentration of bacterial solution to 10 billion / mL; add the equivalent A 0.3% formaldehyde solution of bacterial liquid volume, shake well, and inactivate it in a 37°C thermostat for 24 hours (shaking once every 4h); add Fuchsin staining solution to the bacterial liquid, shake well, and refrigerate at 4°C overnight; Centrifuge at 5000rpm for 10min, discard the supernatant, add physiological saline to suspend the precipitate, after washing 3 times, add physiological saline to the original volume, suspend the prec...

Embodiment 2

[0058] Example 2 Method based on detecting rabbit Klebsiella pneumoniae antibodies

[0059] Preparation of serum to be tested: Collect blood from diseased rabbits or disease-tolerant rabbits, coagulate naturally, separate the serum, put it in a water bath at 56°C for 30 minutes, and set aside.

[0060] Dilution of the serum to be tested: add the diluent to the 96-well microreaction plate, 50μL per well, add 50μL of the serum to be tested in the first well, mix well, and perform 1:2, 1:4, 1:8, 1 respectively :1:16, 1:32, 1:64:, 1:128, 1:256 volume dilution, set positive serum (the positive serum prepared in Example 1), negative serum (the negative serum prepared in Example 1), Diluent control (using the diluent prepared in Example 1).

[0061] Add agglutinogen: add 50μL of detection antigen to each well, shake gently, and place in a refrigerator at 4-8°C for 3 hours.

[0062] Result judgment: Tilt the micro reaction plate about 60°, and observe from the back of the micro reaction plat...

Embodiment 3

[0076] Example 3 Qualitative detection of suspected rabbit Klebsiella pneumoniae

[0077] a. Testing of samples to be tested: 10 strains of suspected Klebsiella pneumoniae were isolated from clinically ill rabbits. The bacteria to be tested were inoculated on TSA agar plates, incubated in a 37°C incubator for 24 hours, and the lawn was washed with physiological saline; Count live bacteria; adjust the concentration of the bacterial solution to 12 billion cells / mL; add formaldehyde according to the amount of the bacterial solution to make the final volume concentration of 0.3%, shake it, and inactivate it in a 37°C incubator for 24 hours (shake once every 4 hours); Add 120μL of Fuchsin staining solution to 1mL of bacterial solution, shake well, and refrigerate overnight at 4°C; centrifuge at 5000rpm for 10min, discard the supernatant, and add physiological saline to suspend the precipitate. After washing 3 times, add physiological saline to the original volume and suspend the precip...

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Abstract

The invention relates to the technical field of organisms, in particular to a rabbit Klebsiella pneumoniae agglutination reaction experimental technique. A rabbit Klebsiella pneumoniae agglutinogen is prepared from rabbit Klebsiella pneumoniae liquid dyed by fuchsin dye liquid. An application method concretely belongs to an application method in agglutination reaction experiments: the rabbit Klebsiella pneumoniae agglutination is combined with agglutinin, and the agglutination result is judged. The rabbit Klebsiella pneumoniae agglutinogen has the advantages that expensive instruments and reagents are not needed, the cost is low, the speed is high, the operation is simple and convenient, the technical requirements on operators are low, the result is intuitional, the rabbit Klebsiella pneumoniae agglutinogen is applied to basic level site detection application, the detection antigen of a rabbit Klebsiella pneumoniae microagglutination reagent kit is dyed, agglutination particles or agglutination blocks are red, the observation is easier, and the detecting method can be used for quantificationally detecting the rabbit serum antibody valence, can also be used for qualitatively detecting the rabbit Klebsiella pneumoniae antigen and is particularly suitable for on-site rabbit Klebsiella pneumoniae diagnosis for basic level veterinary stations and warrens.

Description

Technical field [0001] The present invention relates to the field of biotechnology, in particular to the experimental technology of agglutination reaction of rabbit Klebsiella pneumoniae. Background technique [0002] Klebsiellosis in rabbits is a sporadic disease characterized by pneumonia and purulent inflammation of other organs caused by Klebsiella pneumoniae in rabbits. It is often endemic and rarely causes major diseases. The scale is popular. Rabbits show a series of respiratory symptoms, digestive disorders, miscarriage of female rabbits, severe cases of sepsis and death. Rabbits of various breeds, ages and sexes are all susceptible to this bacteria. The incidence rate is highest in young rabbits before and after weaning and pregnant female rabbits. After infection, the offspring are extremely exhausted and die quickly, which is very serious. Due to the lack of characteristic clinical symptoms and pathological changes of this disease, it is difficult to make an accurate...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/569G01N33/536
Inventor 王孝友杨睿沈克飞徐登峰付利芝黄伟
Owner CHONGQING ACAD OF ANIMAL SCI
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