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Antibody ELISA detection method based on salmonella pullorum X protein coating

A Salmonella and detection method technology, applied in the field of poultry bacterial disease detection, can solve problems such as short maintenance time, achieve the effects of improving detection sensitivity and specificity, and reducing manual judgment errors

Inactive Publication Date: 2021-04-13
JIANGSU LIHUA ANIMAL HUSBANDRY +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Existing studies have shown that the agglutination antibody detected by the agglutination reaction is produced quickly in the body, but the maintenance time is relatively short, and there must be false negative chickens with "agglutination antibody negative and ELISA antibody positive" in the later stage.

Method used

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  • Antibody ELISA detection method based on salmonella pullorum X protein coating
  • Antibody ELISA detection method based on salmonella pullorum X protein coating
  • Antibody ELISA detection method based on salmonella pullorum X protein coating

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Embodiment Construction

[0051] In order to illustrate the technical solution of the present invention more clearly, the technical solution of the present invention is described in further detail below in conjunction with the accompanying drawings:

[0052] Such as figure 1 Described, a kind of antibody ELISA detection method based on X protein coating of Salmonella pullorum, is characterized in that, specific operation steps are as follows:

[0053] Step (1), by preparing and purifying the SPF chicken-derived high-affinity polyclonal antibody of Salmonella pullorum, adopting the co-immunoprecipitation method to screen the dominant antigen X protein;

[0054] Step (2), comparing the dominant antigen X protein obtained by co-immunoprecipitation mass spectrometry;

[0055] Step (3), construction, expression and purification of dominant antigen X protein;

[0056] Step (4), establishment of indirect ELISA method.

[0057] Further, in the step (1), the pulley pulley dominant antigen X protein is an elect...

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Abstract

The invention discloses an antibody ELISA detection method based on salmonella pullorum X protein coating, and relates to the field of poultry bacterial disease detection. An SPF chicken-derived high-affinity polyclonal antibody of salmonella pullorum is prepared and purified, and an electron carrier in an electron transfer chain and a dominant antigen X protein with oxidordeuctase activity are screened and identified by adopting an immunocoprecipitation technology; the exocrine dominant antigen X protein is constructed, expressed and purified by a specific mild method; and an indirect ELISA method for detecting the salmonella pullorum antibody is established. The salmonella pullorum positive antibody can be effectively detected, and the specificity and sensitivity of an existing detection technology are improved; and a powerful tool is provided for purifying salmonella pullorum of enterprises.

Description

technical field [0001] The invention discloses an antibody ELISA detection method based on X protein coating of Salmonella pullorum, and relates to the field of poultry bacterial disease detection. Background technique [0002] Pullorum disease (PD) is a poultry infectious disease caused by Salmonella pullorum (Salmonella pullorum, SP). SP mainly affects chicks aged 2-3 weeks, and is mainly characterized by pullorum, sepsis, myocarditis, and encephalitis, and a large number of deaths will occur. Adult chickens are mostly chronically infected or recessively infected, and vertical transmission causes the infection and death of offspring chicks. In addition, SP host cells are immune cells-macrophages, which lead to severe immunosuppression after infection, so the direct death loss and indirect economic loss caused by PD are relatively huge. At present, developed countries in Europe and the United States have basically achieved the purification of pullorum pullorum. However, i...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/569G01N27/62C07K14/255
CPCC07K14/255G01N33/56916G01N33/6848G01N2333/255G01N2469/20
Inventor 尹丽萍张凯云杨铜戎双琳张瑶牟抑扬刘岳龙张进程立力
Owner JIANGSU LIHUA ANIMAL HUSBANDRY
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