A pan-type inert carrier Escherichia coli and its potential application

An inert carrier, Escherichia coli technology, applied in the field of biomedical technology detection, can solve problems such as non-agglutination, and achieve the effects of perfecting poor specificity, improving poor specificity, huge application value and market prospect

Active Publication Date: 2021-01-05
YANGZHOU UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] In the previous research, the inert carrier Escherichia coli SE1 studied by the applicant only has the function of non-agglutination for chicken serum of different backgrounds within a certain concentration range, but it may have different degrees of agglutination for other animals. Therefore, the inert carrier Escherichia coli SE1 Bacillus SE1 can only be used to develop chicken agglutination assay and its application, and its application has certain limitations

Method used

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  • A pan-type inert carrier Escherichia coli and its potential application
  • A pan-type inert carrier Escherichia coli and its potential application
  • A pan-type inert carrier Escherichia coli and its potential application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0039] Example 1 Acquisition and verification of pan-type inert carrier E. coli SE1H

[0040] Inoculate the inert carrier Escherichia coli SE1 (preservation number is CGMCC No.17339) in the LB liquid medium, shake at 37°C for 12 hours, draw 30 μL of the bacterial liquid into the LB solid medium for streak culture, and culture at 37°C for 16-18 hours Obtain the second-generation colony, pick the second-generation single colony and inoculate it in LB liquid medium, and use LB liquid and solid medium as a cycle to cultivate alternately. Starting from the 40th generation, the single colony obtained is a pan-type inert carrier bacterium SE1H, ​​in fact, through the 41st to 60th generations of continuous subculture, any generation of them also has the characteristics of the pan-type inert carrier bacteria SE1H.

[0041] Take 1 mL of the SE1H cultured overnight to prepare DNA template by boiling method. The Escherichia coli β-glucuronidase gene uidA was amplified by PCR, identified ...

Embodiment 2

[0049] Example 2 Test of no non-specific agglutination phenomenon of pan-type inert carrier Escherichia coli SE1H and human and animal source serum and whole blood of different backgrounds

[0050] Centrifuge the Escherichia coli SE1H bacterial solution obtained from the overnight culture in Example 1 at 4° C. at 4000 rpm for 10 min, discard the supernatant, resuspend the bacteria sludge with sterile saline, wash by centrifugation three times, and then resuspend to a concentration gradient of different concentrations of bacteria. Before the test, the bacterial liquid was mixed with a vortex instrument, and the agglutination test was performed with sterile saline and SPF chicken serum to ensure that the test bacterial liquid had no self-agglutination and no non-specific agglutination. In the ultra-clean bench (20 ° C ~ 25 ° C), take several pieces of ordinary glass plates with clean surfaces, and use sterile pre-cooled PBS to 4 ° C to resuspend and wash the carrier bacteria for ...

Embodiment 3

[0056] Example 3 Test and verification of the surface expression of pan-type inert carrier bacteria Escherichia coli SE1H and the ability to carry the avian-derived Salmonella antigenic factor P

[0057] (1) Amplification of gene p encoding antigenic factor P expressed by avian Salmonella

[0058] According to the whole genome sequence of Salmonella pullorum ATCC 9120 strain in NCBI GenBank (NCBI accession number: CP012347.1), the whole genome sequence of Salmonella pullorum S44987_1 strain (NCBI accession number: LK931482.1), the whole genome sequence of Salmonella pullorum S06004 strain ( NCBI accession number: CP006575.1), whole genome sequence of Salmonella pullorum QJ-2D-Sal strain (NCBI accession number: CP022963.1), whole genome sequence of Salmonella gallinarum typhi 287 / 91 strain (NCBI accession number: AM933173.1) , Salmonella gallinarum typhi 9184 strain genome sequence (NCBI accession number: CP019035.1) published full-length genome sequence, search and compare the...

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Abstract

The invention discloses a pan-type inert carrier Salmonella S9H and its potential application. The pan-type inert carrier Salmonella S9H is obtained from the inert carrier S9 using LB solid and liquid medium for continuous in vitro culture and subculture to the 40th generation. It can be compared with human, mouse, bovine, and pig at the working concentration of bacteria. Non-specific agglutination reaction does not occur in serum and whole blood of source and poultry (including chicken, duck, goose, turkey, pigeon, quail), and has carrying and surface expression displaying human, mouse, bovine and porcine sources The characteristics of different antigenic factors from poultry sources (including chicken, duck, goose, turkey, pigeon, quail) can be applied to the development, improvement and It improves the technical bottleneck of poor specificity and poor sensitivity of the agglutination test of the existing agglutination antigen antibody detection, and has wide application value and market prospect.

Description

technical field [0001] The invention belongs to the field of biomedical technology detection, and specifically relates to a pan-type inert carrier Escherichia coli and its potential application. Pig and poultry (including chicken, duck, goose, turkey, pigeon, quail) with different genetic backgrounds and various animal serum and whole blood do not have non-specific agglutination reactions. Background technique [0002] In the diagnosis, monitoring and prevention and control of livestock and poultry diseases, currently commonly used diagnostic techniques include serological diagnostic techniques and pathogenic detection techniques. In the diagnosis of bacterial diseases, although the isolation and identification of bacteria is the gold standard method for diagnosing bacterial infections, there are deficiencies such as long culture period, limited sampling site, limited sampling time, low detection rate, cumbersome and complicated operation, etc. If the isolation and identifi...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/21C12N15/70G01N33/569C12R1/19
CPCC12N15/70C07K14/255C07K14/245G01N33/56916C12N1/205C12N1/36G01N2333/245
Inventor 朱国强杨斌羊扬孟霞夏芃芃段强德王亨朱晓芳
Owner YANGZHOU UNIV
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