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Generic inert carrier salmonella and potential application thereof

An inert carrier, the technology of Salmonella, applied in the field of biomedical technology detection, can solve the problems of affecting and affecting the effect of epidemic disease purification, epidemic disease purification, interference detection and diagnosis results, etc., to achieve perfect specificity, improve specificity, huge The effect of application value and market prospect

Active Publication Date: 2020-08-07
YANGZHOU UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

And it is worth noting that, in view of the fact that the working concentration of the agglutinated antigen needs to contain a higher concentration of bacteria and lead to non-specific cross-reaction effects, this non-specific cross-reaction disadvantage will inevitably affect or even significantly interfere with the detection and diagnosis results, thereby seriously affecting the epidemic. Purification effect and advancement of disease purification work process
[0005] In the previous research, the inert carrier bacterium S9 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, its inert carrier bacterium S9 Bacteria can only be used to develop chicken agglutination test and its application, and its application has certain limitations

Method used

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  • Generic inert carrier salmonella and potential application thereof
  • Generic inert carrier salmonella and potential application thereof
  • Generic inert carrier salmonella and potential application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] Example 1 The acquisition and verification of the pan-type inert carrier bacteria S9H

[0042] The inert carrier Salmonella S9 (preservation number is CGMCCNo.17340) was inoculated in the LB liquid medium, shaken at 37°C for 12 hours, then sucked 30 μL of the bacterial liquid into the LB solid medium for streak culture, and cultured at 37°C for 16-18 hours to obtain the first For the second-generation colony, pick the second-generation single colony and inoculate it in LB liquid medium. According to the same conditions as above, use LB liquid and solid medium as a cycle to cultivate alternately. Starting from the 40th generation, the single colony obtained is pantropic. Type inert carrier bacteria S9H, in fact, continues to be passed down forty-first to sixty generations, and any generation of them also has the above-mentioned characteristics of pan-type inert carrier bacteria S9H.

[0043] Use the primers of the fimW gene of Salmonella species reported by the applicant...

Embodiment 2

[0052]Example 2 Carrier bacterium Salmonella S9H and human and animal source serum and whole blood of different backgrounds without non-specific agglutination

[0053] According to the method of Example 1, the carrier bacterium S9 was alternately subcultured with LB agar and LB liquid medium for 40 generations to obtain the pan-type carrier bacterium Salmonella S9H, and the bacterial liquid was centrifuged at 4°C 4000rpm for 10min, and the supernatant was discarded. Resuspended in saline, centrifuged and washed three times, and then resuspended 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, an...

Embodiment 3

[0060] Example 3 Test and verification of the surface expression of the carrier bacterium Salmonella S9H and the ability to carry the avian-derived Salmonella antigenic factor P

[0061] (1) Amplification of gene p encoding antigenic factor P expressed by avian Salmonella

[0062] 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 full-length frag...

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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, the characteristics of carrying and surface expression displaying human sources, mouse sources, cattle sources, pig sources and poultry sources (including chickens, ducks, gooses, turkeys, pigeonsand 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, and has a 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 Salmonella and its potential application. And poultry (including chicken, duck, goose, turkey, pigeon, quail) different genetic background, human and various animal serum and whole blood do not have non-specific agglutination. Background technique [0002] In disease prevention and control and epidemiological research, serological detection techniques are often used to diagnose whether animals are infected or carry a specific pathogen, among which agglutination test is a classic serological rapid diagnosis widely used in medicine and veterinary clinics. method. The principle of agglutination test is that bacterial granular antigens will agglutinate and coagulate within a few minutes after combining with corresponding serum antibodies in the presence of electrolytes and at a suitable temperature, forming agglomerated small pieces ...

Claims

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Application Information

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IPC IPC(8): C12N1/20G01N33/569C12R1/42
CPCC12N1/20G01N33/56916C12R2001/42C12N1/205Y02A50/30G01N2469/20G01N2333/245G01N2333/255C07K14/255C07K14/245
Inventor 朱国强杨斌羊扬孟霞夏芃芃段强德朱晓芳
Owner YANGZHOU UNIV
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