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Generic inert carrier escherichia coli and potential application thereof

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: 2020-08-21
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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  • Generic inert carrier escherichia coli and potential application thereof
  • Generic inert carrier escherichia coli and potential application thereof
  • Generic inert carrier escherichia coli and potential application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0039] Example 1 Obtaining and verifying the ubiquitous inert vector Escherichia coli SE1H

[0040] The inert carrier Escherichia coli SE1 (preservation number CGMCC No.17339) was inoculated into LB liquid medium and shaken at 37°C for 12 hours. Then 30μL of bacterial liquid was drawn and streaked in LB solid medium, and cultured at 37°C for 16-18 hours. Obtain the second generation of colonies, pick a single colony of the second generation and inoculate it in LB liquid medium, use LB liquid and solid medium as a cycle to alternate culture. Starting from the 40th generation, the single colony obtained is a ubiquitous inert carrier bacteria SE1H, ​​in fact, by continuing to pass down from the 41st to the 60th generation, any of them will also have the characteristics of the ubiquitous inert carrier bacteria SE1H.

[0041] Take 1 mL of the above-mentioned overnight cultured SE1H bacterial liquid and prepare DNA template by boiling method. The E. coli β-glucuronidase gene uidA was am...

Embodiment 2

[0049] Example 2 Test for non-specific agglutination of pan-type inert carrier Escherichia coli SE1H with human and animal serum and whole blood of different backgrounds

[0050] The Escherichia coli SE1H bacterial solution obtained by overnight culture in Example 1 was centrifuged at 4°C at 4000 rpm for 10 minutes, the supernatant was discarded, and the bacterial sludge was resuspended in sterile normal saline, centrifuged and washed three times, and then resuspended to a concentration gradient of different concentrations of bacteria. Before the test, mix the bacterial liquid with a vortexer, and perform agglutination test with sterile normal saline and SPF chicken serum to ensure that the test bacterial liquid does not self-agglomerate or non-specific agglutination. Take several common glass plates with clean surfaces in an ultra-clean bench (20℃~25℃), centrifuge and resuspend the carrier bacteria with sterile pre-cooled PBS to 4℃ for 3 times, then resuspend and dilute to the sp...

Embodiment 3

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

[0057] (1) Amplification of the coding gene p of Avian Salmonella expressing antigen factor P

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

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Abstract

The invention discloses generic inert carrier salmonella S9H and a potential application thereof. The generic inert carrier salmonella S9H is obtained by continuously culturing inert carrier bacteriaS9 in vitro by using an LB solid and liquid culture medium for passage to the fortieth generation, under the condition of the quantity of bacteria with working concentration, the non-specific agglutination reaction with serum and whole blood of human sources, mouse sources, cattle sources, pig sources and poultry sources (including chickens, ducks, gooses, turkeys, pigeons and quails) can be avoided; due to the characteristics of carrying and surface expression displaying human sources, mouse sources, cattle sources, pig sources and poultry sources (including chickens, ducks, gooses, turkeys,pigeons and quails) for different resistance factors are realized, the generic inert carrier salmonella S9H can be applied to development of an indirect agglutination test detection method for simply,conveniently and quickly detecting human and various animal antigens or infected antibodies, improves and perfects the technical bottleneck of poor specificity and sensitivity of an existing agglutination test for agglutination antigen and antibody detection. The product 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 applications. The pan-type inert carrier Escherichia coli can interact with human origin, mouse origin, bovine origin, and human origin under the working concentration of bacteria. The serum and whole blood of pig and poultry sources (including chickens, ducks, geese, turkeys, pigeons, quail) of different genetic backgrounds 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 diagnosis techniques and pathogen detection techniques. In the diagnosis of bacterial diseases, although the isolation and identification of bacteria is the gold standard method for the diagnosis of bacterial infections, it has the disadvantages o...

Claims

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Application Information

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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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