Pullorum agar gel precipitating antigen as well as preparation method and application thereof

A pullorum and antigen technology, which is applied to biochemical equipment and methods, microorganism-based methods, instruments, etc., can solve the problems of insufficient clarity of agglutination images, prolonged purification time, low sensitivity, etc., and achieves simple and reasonable process methods. Purification cycle, clear effect of agglutination image

Active Publication Date: 2015-07-22
JIANGSU INST OF POULTRY SCI
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  • Abstract
  • Description
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AI Technical Summary

Problems solved by technology

However, in actual production, there are still defects in the use of pullorum agglutinated antigen for quarantine and purification. For example, individuals who are weakly positive in the agglutinated antigen test result still need to be quarantined again after an interval of 2-3 weeks, which not only increases the cost of feeding, but also prolongs the time. The purification time is shortened, and the risk of horizontal transmission of the disease is also increased; since Salmonella pullorum infection of young chickens cannot reproduce in large quantities in the body, it usually does not produce high levels of agglutinated antibodies, resulting in the detection

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  • Pullorum agar gel precipitating antigen as well as preparation method and application thereof
  • Pullorum agar gel precipitating antigen as well as preparation method and application thereof
  • Pullorum agar gel precipitating antigen as well as preparation method and application thereof

Examples

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Effect test

Embodiment 1

[0020] 1) Separation, purification and staining microscopy

[0021] Salmonella pullorum sp8441 strain was isolated from the oviduct of 6-month-old white legion laying hens in a chicken farm in Beijing in December 1984. The specific separation method is as follows: aseptically cut a small section of the oviduct and add it to nutrient broth for overnight culture at 37°C, then inoculate it on Martin agar medium, and culture it at 37°C for 24 hours. Pick a colony with a diameter of 1.0mm-2.0mm, off-white transparent, and smooth surface, inoculate it on Martin agar medium, and incubate at 37°C for 24 hours. The purely cultured bacteria were subjected to Gram staining microscope inspection, and then observed with an optical microscope, showing elongated bacilli with slightly rounded ends.

[0022] Salmonella pullorum SP9905 strain was isolated from the cecum contents of 11-month-old Langshan chickens in a chicken farm in Yangzhou City, Jiangsu Province in May 1999. The specific se...

Embodiment 2

[0042] Salmonella pullorum SP8441 and SP9905 strains were resuscitated and multiplied in liquid medium respectively, inoculated on solid plate medium, cultured at 37°C for 24 hours, and typical colonies were selected to prepare seed bacterial liquid. Inoculate the solid flat bottle culture medium with the seed bacterial liquid at 2.0% of the medium volume, incubate at 37°C for 24 hours, wash the bacterial lawn with sterile physiological saline, collect the bacterial liquid after filtering through sterilized gauze, and centrifuge at 4000r / min for 15 minutes Remove the supernatant, then re-suspend the bacterial sludge with sterile physiological saline, and adjust the bacterial concentration of SP8441 and SP9905 to 1.0×10 per ml by adjusting the turbidimetric method. 10 CFU, mixed in a volume ratio of 4:6. After freezing and thawing the mixed bacterial solution once, mix it fully with an equal volume of bacterial lysis buffer, act at 4°C for 1 hour, centrifuge at 12,000 r / min for...

Embodiment 3

[0049] According to the preparation method described in Example 2, the agar-enhanced antigen of pullorum agar was prepared, and the whole blood of 8-13-week-old Suqin yellow chicken, Beijing powder layer, Hailan layer, green-shell layer, and Xianju chicken was carried out. Agar diffusion test and commercialized pullorum agglutination test antigen (purchased from Qingdao Yibang Bioengineering Co., Ltd.) were used to conduct whole blood plate agglutination test, and all chickens were autopsied for bacterial isolation. The results are shown in Table 4.

[0050] It can be seen from Table 4 that among the 492 young chickens tested, 60 chickens were positive for Qiong Kuo antigen, and the bacterial isolation rate was 100%, which was significantly higher than the positive detection rate of agglutinating antigen (P<0.05). The above-mentioned results show that the Qiongjian antigen prepared by the method of the present invention has a good detection effect on young chickens of different...

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Abstract

The invention discloses a pullorum agar gel precipitating antigen as well as a preparation method and an application thereof, and belongs to the technical field of veterinarian diagnosis. Salmonella pullorum SP8441 and salmonella pullorum SP9905 are subjected to recovery and passage respectively to form a seed bacterium liquid, then the seed bacterium liquid is inoculated with a solid culture medium for proliferation, after a culture solution is removed centrifugally, bacterial protein is extracted with a lysis buffer, and the pullorum agar gel precipitating antigen is prepared. The technological method is simple, reasonable, scientific, stable in production and low in cost, the prepared pullorum agar gel precipitating antigen product has the advantages of high sensitivity and high specificity, the pullorum purification process can be simplified, the detection application range can be enlarged, and the agar gel precipitating antigen is relatively ideal in pullorum detection.

Description

technical field [0001] The invention belongs to the technical field of veterinary diagnosis, and in particular relates to a chicken pullorum agar expansion antigen and a preparation method and application thereof. Background technique [0002] Pullorum is the most serious bacterial infectious disease in poultry farming. Its pathogen is Salmonella pullorum. The disease can be transmitted vertically through eggs, and can also be transmitted horizontally through respiratory tract, digestive tract, conjunctiva and damaged skin wounds. After Salmonella pullorum infection, it can survive in the host body for a long time until it colonizes the ovary, fallopian tube, testis and other sexual organs after sexual maturity, and produces a high level of agglutination antibody continuously. Whole blood (serum) and elimination of positive chickens can cut off the vertical transmission of pathogens. However, in actual production, there are still defects in the use of pullorum agglutinated ...

Claims

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Application Information

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IPC IPC(8): C12N1/20C12N1/06G01N33/569C12R1/42
CPCC12N1/20C12N1/205C12R2001/42G01N33/569
Inventor 龚建森顾蓓蓓许明苗晋锋张笛朱春红窦新红刘学贤徐步童海兵邹剑敏
Owner JIANGSU INST OF POULTRY SCI
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