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Lawsonia intracellularis flgE recombinant protein and lawsonia intracellularis antibody detection kit

A technology of recombinant protein and Lawsonia, applied in biological testing, bacterial peptides, measuring devices, etc., to achieve high sensitivity, high specificity, and rapid detection

Active Publication Date: 2021-06-11
湖南康保特生物科技有限公司 +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although many domestic studies have shown that the ELISA antibody detection method based on LI recombinant outer membrane or surface protein has good specificity and sensitivity, but there is no mature commercial LI antibody detection kit in China at present.

Method used

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  • Lawsonia intracellularis flgE recombinant protein and lawsonia intracellularis antibody detection kit
  • Lawsonia intracellularis flgE recombinant protein and lawsonia intracellularis antibody detection kit
  • Lawsonia intracellularis flgE recombinant protein and lawsonia intracellularis antibody detection kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0011] Example 1 Preparation of Lawsonia intracellulare flgE recombinant protein

[0012] According to the gene sequence of the flagellar hook matrix complex protein flgE in the LI complete gene sequence (accession numbers: NC_008011.1 and CP004029.1) in GenBank, PrimerSelect was used to design primers, and the corresponding enzyme cutting sites EcoRI, SalⅠand protective bases, upstream primer fP: 5'-CCGGAATTCATGAGTCTTACAGCAGGAATG-3'(EcoRⅠ)(SEQ ID NO.3); downstream primer rP: 5'-ACGCGTCGACACGCTTCATATTTACAACTGT-3'(SalⅠ)(SEQ ID NO.4 ).

[0013] According to the instructions of the bacterial DNA extraction kit, the DNA in the feces of LI-infected pigs was extracted and used as a template. HS DNA Polymerase system carries out PCR amplification (PCR amplification system is: HSDNA Polymerase: 1μL; dNTP: 4μL; Buffer: 10μL; dH 2 0: 29 μL; template: 2 μL; upstream and downstream primers: 2 μL each. PCR amplification conditions are: pre-denaturation at 98°C for 2min; denaturation:...

Embodiment 3

[0032] Embodiment 3 Indirect ELISA method of the present invention detects Lawsonia intracellulare antibody

[0033] Adopt the indirect ELISA method of the present invention to detect 2 sow farm pigs (A farm and B farm) different parity sow serum, the results are shown in the following table 2, showing the low parity to high parity sow serum in 2 pig farms The antibody levels decreased in turn and the antibody tests were all positive (in pig farms with standardized feeding management, pigs infected with LI can obtain strong immune protection).

[0034] Table 2 Antibody detection results of different parity sow serum

[0035]

[0036] ②Adopt the indirect ELISA method of the present invention to detect commercial pig serum in different stages of 3 pig farms (C farm, D farm and E farm). The primiparous sows and multiparous sows in Farm C were raised separately, and the serum came from the pigs of the multiparous sows. Farms D and E were commercial pig farms raised on traditio...

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Abstract

The amino acid sequence of the lawsonia intracellularis flgE recombinant protein is as shown in SEQ ID NO. 1. The kit prepared from the lawsonia intracellularis flgE recombinant protein is high in specificity and good in sensitivity and repeatability, can be used for detecting lawsonia intracellularis antibodies and discovering infection of swinery in the early stage, is high in detection speed, can be used for evaluating the LI infection state of the swinery, and is convenient for guiding protection, prevention and control of infection of the lawsonia intracellularis.

Description

technical field [0001] The invention belongs to the field of veterinary biotechnology, relates to Lawsonia intracellulare flgE recombinant protein, and also relates to a kit for establishing an indirect ELISA (flgE-ELISA) method for detecting Lawsonia intracellulare antibodies by using the flgE recombinant protein as an antigen. It is used for the detection of Lawsonia intracellulare flgE protein antibody, so as to judge the infection of Lawsonia intracellulare in pigs and monitor the prevalence of the disease. Background technique [0002] Porcine proliferative enteropathy (PPE) is an infectious intestinal disease characterized by proliferation of intestinal epithelial cells and thickening of intestinal mucosa caused by Lawsonia intracellularis (LI). It is one of the common diseases in chemical farms. The harm to the pig industry is not only the occasional acute death of medium and large pigs. caused significant economic losses. Since feed manufacturers stopped producing ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/195C12N15/31G01N33/569G01N33/68
CPCC07K14/195G01N33/56911G01N33/6854G01N2333/195G01N2469/20Y02A50/30
Inventor 余兴龙廖荣莉郑金赵墩
Owner 湖南康保特生物科技有限公司
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