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A kind of pullorum agglutinating antigen and preparation method thereof

A pullorum and antigen technology, which is applied to biochemical equipment and methods, microorganism-based methods, instruments, etc., can solve the problems of low sensitivity of dyeing antigens, slow diagnosis speed, and inability to achieve complete purification of breeder flocks, and achieves a technological The method is scientific and reasonable, easy to observe, and the effect of low cost

Active Publication Date: 2020-06-23
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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  • A kind of pullorum agglutinating antigen and preparation method thereof
  • A kind of pullorum agglutinating antigen and preparation method thereof
  • A kind of pullorum agglutinating antigen and preparation method thereof

Examples

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preparation example Construction

[0059] B: the preparation method of pullorum agglutinating antigen

[0060] (1) Cultivate bacteria: Pass the resuscitated strains on TG agar for 2 to 3 generations continuously, pick enough colonies to spread evenly on the whole plate on TG agar, and culture at 37°C for 18 to 24 hours.

[0061] (2) Bacteria collection: Wash the bacterial cells with a sufficient amount of 0.2% normal saline formalin, centrifuge at 4000 rpm for 40 min, collect bacterial precipitates, and resuspend.

[0062] (3) Inactivation: Add a sufficient amount of 0.4% formalin saline, resuspend, and let stand at 4°C for 24 hours. Then use an inoculation loop to dip in the bacterial solution and draw a line on the MacConkey plate for sterility testing.

[0063] (4) Alcohol precipitation: After passing the sterility test, centrifuge at 4000rpm for 40min, discard the supernatant, add twice the volume of absolute ethanol, resuspend, and stand at 4°C for 24-36h until the bacterial precipitation is complete, rem...

Embodiment 1

[0077] 100 naturally infected chicken sera provided by company A were tested with self-made antigens, and commercialized pullorum chicken typhoid polyvalent stained plate agglutinated antigen (purchased from China Veterinary Drug Control Institute, batch number 201402, referred to as CVCC antigen) was used as the control antigen. The results were judged according to the judgment standard of the glass plate agglutination test. The results showed that the number of strong positives for self-made antigens and the antigens of the Institute of Prisons were 47 and 40 respectively; the total positives were 90 and 88 respectively. The total positive rate of the self-made antigen was 2% higher than that of the CVCC antigen, and the strong positive rate was 7% higher than that of the CVCC antigen (Table 5, Table 6).

[0078] Table 5. The results of the self-made antigen and the antigen detection of A company's naturally infected chicken serum

[0079]

[0080]

Embodiment 2

[0082] Two groups of naturally infected chicken sera provided by companies B and C were tested with self-made antigens, and commercialized pullorum polyvalent stained plate agglutination antigen (purchased from China Veterinary Drug Control Institute, batch number 201402) was used as the control antigen. The results were judged according to the judgment standard of the glass plate agglutination test. The results showed that the self-made antigens were 8.43% and 33.33% higher than the Institute of Prisons antigens in terms of total positive rate. In terms of strong positive rates, the self-made antigens were 7.36% and 19.29% higher than those of the Institute of Prisons, respectively (Table 6). The above results show that the detection effect of the antigen prepared by the method of the present invention is obviously better than that of the commercialized antigen, which is beneficial to improving the quarantine quality of pullorum pullorum.

[0083] Table 6 The positive rate o...

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Abstract

The invention relates to an agglutinated antigen of pullorum pullorum and a preparation method thereof, mainly involving bacterial strains of Salmonella pullorum standard strain C79‑1 and wild isolate S44 after screening. The wild isolate of Salmonella pullorum S44, classified and named: Salmonella enterica subsp. enterica pullorum, Latin name: Salmonella enterica subsp.enterica pullorum, was screened from Salmonella pullorum isolated from chickens, and preserved in China General Microbiology Center of Microbial Culture Collection Management Committee, No. CGMCC No.14258. The method of the present invention is scientific and reasonable, stable in production and low in cost, and the selected pullorum stained agglutination antigen production strain has good antigenicity and low variation rate, and the produced pulley pullorum stained agglutination antigen product has high sensitivity, strong specificity, and rapid diagnosis , The agglutination effect is easy to observe and so on.

Description

technical field [0001] The invention relates to an agglutinated antigen of pullorum pullorum and a preparation method thereof, mainly involving bacterial strains of Salmonella pullorum standard strain C79-1 and wild isolate S44 obtained through separation and screening. Background technique [0002] Pullorum pullorum is caused by Salmonella pullorum pullorum. It is an acute or chronic infectious disease that chickens of all ages can infect. The disease can spread both horizontally and vertically, posing a huge threat to the development of poultry farming. Diseased chicks have a high morbidity and mortality rate and 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)" puts forward assessment requirements for the purification of pullorum pullorum. In 2020, the Salmonellosis of ...

Claims

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Application Information

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Patent Type & Authority Patents(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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