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Proteus mirabilis antigen identification and application thereof in detection of proteus infection

A technology of Proteus mirabilis and Proteus vulgaris, applied in the field of biochemistry, can solve the problems of lack of diagnostic antigen evaluation, lack of detection or diagnostic methods, etc., and achieve the effect of strong antigenicity

Pending Publication Date: 2021-09-07
CHONGQING ACAD OF ANIMAL SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

In terms of serological detection methods, one is the lack of evaluation of diagnostic antigens, and the other is the lack of corresponding applicable detection or diagnostic techniques.

Method used

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  • Proteus mirabilis antigen identification and application thereof in detection of proteus infection
  • Proteus mirabilis antigen identification and application thereof in detection of proteus infection
  • Proteus mirabilis antigen identification and application thereof in detection of proteus infection

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Embodiment 1

[0039] Embodiment 1 Operation method

[0040] Strains: Proteus mirabilis, V. vulgaris, Escherichia coli, Staphylococcus aureus, Cryptobacterium pyogenes, and Corynebacterium pseudotuberculosis were preserved and provided by the Veterinary Research Institute of Chongqing Academy of Animal Sciences.

[0041] Main reagents: Bacterial membrane protein separation kit was purchased from Bebo; alkaline phosphatase-labeled goat anti-mouse IgG antibody and horseradish peroxidase (HRP)-labeled goat anti-mouse IgG antibody were purchased from EarthOx Life Sciences ; Ni-NTA filler was purchased from Beijing Nuohe Biotechnology Co., Ltd.; Quick Start Bradford was purchased from Bio-Rad Life Medical Products (Shanghai) Co., Ltd.; BCTP / NBT was purchased from Shanghai Sangon Bioengineering Co., Ltd.; TMB single-component chromogenic solution Bought from Suo Laibao Biotechnology Co., Ltd.; ELISA plate was purchased from Corning Life Science (Wujiang) Co., Ltd.; gelatin was purchased from Guang...

Embodiment 2

[0052] Embodiment 2 experimental result

[0053] 1. Membrane protein extraction and antiserum recognition reaction: Membrane protein was extracted from freshly cultured Proteus mirabilis cells using a bacterial membrane protein extraction kit, and SDS-PAGE showed that the extracted protein was as follows: figure 1 shown. Western blotting showed that the extracted protein could react with the antiserum of Proteus mirabilis challenged mice, presenting multiple bands with relative molecular weights ranging from 75ku to 25ku, such as figure 2 shown.

[0054] 2. Mass spectrometry identification in sequence analysis: Excise the corresponding protein band on the SDS-PAGE gel for LC-MS / MS analysis to obtain 4 protein sequences, as shown in SEQ ID NO.1-SEQ ID NO.4 The sequence was searched by BLAST, and the above four sequences were respectively related to the outer membrane porin (OMP), serine hydrolase (SH), glyceraldehyde-3-phosphate dehydrogenase (glyceraldehyde) of Proteus -3-...

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Abstract

The invention relates to the field of biochemistry, and particularly relates to proteus mirabilis antigen identification and application thereof in detection of proteus infection. The invention provides proteus antigen identification and preparation and feasibility of an antigen in an indirect ELISA method for detecting an anti-proteus IgG antibody. The indirect ELISA method can effectively distinguish proteus mirabilis infection, proteus vulgaris infection, escherichia coli infection, staphylococcus aureus infection, cryptobacter pyogenes infection and corynebacterium pseudotuberculosis infection, and which indicates that the antigen can be used for detecting and diagnosing proteus mirabilis infection and proteus vulgaris infection.

Description

technical field [0001] The invention relates to the field of biochemistry, in particular to the identification of Proteus mirabilis antigen and its application in detecting Proteus infection. Background technique [0002] Proteus belongs to the Enterobacteriaceae Proteus genus, Gram-negative bacteria. The bacteria are widely distributed in nature, mostly in polluted water, soil, air and human or animal excreta. Relevant foreign data show that Proteus can be divided into five named species, namely Proteus vulgaris, Proteus mirabilis, Proteus myxofaciens, Proteus penneri ) and Proteus hauseri and 3 unnamed species. According to domestic data, the Proteus genus is divided into four types: P. vulgaris, P. mirabilis, P. myxofaciens and P. penneri. Proteus mirabilis and Proteus vulgaris are common opportunistic and foodborne pathogens. [0003] The antigens of Gram-negative bacteria mainly include O antigen, outer membrane protein, flagella, pili, capsule and so on. Proteus c...

Claims

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Application Information

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IPC IPC(8): C07K14/24C12N15/70C12N1/21G01N33/569G01N33/543G01N33/58G01N33/68C12R1/19
CPCC07K14/24C12N15/70G01N33/56916G01N33/543G01N33/581G01N33/6854C12N2800/22G01N2333/24G01N2469/10Y02A50/30
Inventor 沈克飞付利芝杨睿张素辉徐登峰郑华
Owner CHONGQING ACAD OF ANIMAL SCI
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