Pullorum disease agglutination antigen and preparation method thereof

A chicken pullorum and antigen technology, applied in biochemical equipment and methods, methods based on microorganisms, instruments, etc., can solve the problems of low sensitivity of dyed antigens, inability to achieve thorough purification of breeder flocks, and slow diagnosis speed, etc. The method is scientific and reasonable, easy to observe, and the effect of rapid diagnosis

Active Publication Date: 2017-12-08
YANGZHOU UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] The existing commercialized "Polyella pullorum polyvalent staining plate agglutination antigen" is prepared from the standard strain C79-1 and the mutant strain C79-7 of Salmonella pullorum isolated in the early years, but in recent years it has been used in practical applications. It was found that the dyed antigen had defects such as low sensitivity and slow diagnosis speed, which resulted in the inability of the tested breeder flock to achieve the goal of thorough purification

Method used

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  • Pullorum disease agglutination antigen and preparation method thereof
  • Pullorum disease agglutination antigen and preparation method thereof
  • Pullorum disease agglutination antigen and preparation method thereof

Examples

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preparation example Construction

[0059] B: Preparation method of pullorum agglutination antigen

[0060] (1) Cultivation of bacteria: Pass the resuscitated strains on TG agar for 2 to 3 consecutive generations, pick enough colonies on TG agar to coat the whole plate uniformly, and incubate at 37°C for 18-24 hours.

[0061] (2) Collect bacteria: Wash the bacteria with a sufficient amount of 0.2% formalin saline, centrifuge at 4000 rpm for 40 minutes, collect the bacterial pellet, and resuspend.

[0062] (3) Inactivation: Add a sufficient amount of 0.4% formalin saline, resuspend it, and leave it at 4°C for 24 hours. Then use the inoculating loop to dip the bacterial solution to streak on the MacConkey plate for sterility test.

[0063] (4) Alcohol precipitation: After passing the sterility test, centrifuge at 4000 rpm for 40 minutes, discard the supernatant, add twice the volume of absolute ethanol, resuspend, and let stand at 4°C for 24 to 36 hours until the bacterial precipitation is complete, remove the upper layer...

Embodiment 1

[0077] The self-made antigen detection company A provided 100 samples of naturally infected chicken sera, and at the same time commercialized pullorella chicken typhoid multivalent staining plate agglutination antigen (purchased from China Veterinary Drug Inspection Institute, batch number 201402, referred to as CVCC antigen) as the control antigen. The result is judged according to the judgment standard of glass plate agglutination test. The results showed that the number of strong positives for the self-made antigen and the Central Prison Institute antigen were 47 and 40 respectively; the total positive numbers were 90 and 88 respectively. The total positive rate of self-made antigen is 2% higher than that of CVCC antigen, and the strong positive rate is 7% higher than that of CVCC antigen (Table 5, Table 6).

[0078] Table 5. Self-made antigens and the results of the detection of the serum from the naturally infected chickens of A company

[0079]

[0080]

Embodiment 2

[0082] The self-made antigen was used to detect the two groups of naturally infected chicken sera provided by Company B and Company C. At the same time, commercial pullorum typhoid multivalent staining plate agglutination antigen (purchased from China Veterinary Drug Administration, batch number 201402) was used as the control antigen. The result is judged according to the judgment standard of glass plate agglutination test. The results showed that in terms of the total positive rate, the self-made antigen was 8.43% and 33.33% higher than the Central Prison antigen respectively. In terms of the strong positive rate, the self-made antigens were 7.36% and 19.29% higher than the Central Prison antigen respectively (Table 6). The above results show that the detection effect of the antigen prepared by the method of the present invention is significantly better than that of the commercial antigen, which is beneficial to improve the quarantine quality of pullorum.

[0083] Table 6 The ...

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Abstract

The invention relates to a pullorum disease agglutination antigen and a preparation method thereof and mainly relates to a bacterial strain which is salmonella pullorum standard strain C79-1 and a screened wild isolated strain S44. The wild isolated strain S44 of salmonella pullorum is classified and named as follows: salmonella enterica intestinal sub-salmonella pullorum, of which the Latin name is Salmonella enterica subsp. enterica pullorum, which is screened from the salmonella pullorum separated from chicken and which is collected at China General Microbiological Culture Collection Center with the collection No.: CGMCC No. 14258. The method provided by the invention is scientific and reasonable, stable in production and low in cost; the selected pullorum disease agglutination antigen production strain is high in antigenicity and low in aberration rate; and the prepared pullorum disease agglutination antigen product has the advantages of high sensibility, high specificity, quick diagnosis, easiness in observing agglutination effect, and the like.

Description

Technical field [0001] The present invention relates to a pullorum pullorum agglutination antigen and a preparation method thereof. The bacterial strains mainly involved are Salmonella pullorum standard strain C79-1 and a wild isolate S44 obtained by separation and screening. Background technique [0002] Pullorum pullorum is caused by Salmonella pullorum. It is an acute or chronic infectious disease that can be infected by chickens of all ages. The disease can be transmitted horizontally and vertically, posing a huge threat to the development of poultry breeding. The high morbidity and fatality rate of sick chicks can also cause a decline in the performance of adult chickens. In my country and other developing countries, the pollution of Salmonella pullorum is quite serious. The "National Medium and Long-term Animal Disease Prevention and Control Plan (2012-2020)" put forward assessment requirements for the purification of pullorum. In 2020, all breeder farms across the country...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20G01N33/569C12R1/42
CPCC12N1/20C12N1/205C12R2001/42G01N33/56911G01N2333/255
Inventor 陈素娟彭大新秦涛张伟伟
Owner YANGZHOU UNIV
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