146 results about "Salmonella diarizonae" patented technology

The invention discloses a method for simultaneously and quickly detecting salmonella typhimurium, escherichia coli O157:H7 and listeria monocytogenesis in food. The method comprises the following steps of: preparing nano immunomagnetic particles of the targeted typhimurium, escherichia coli O157:H7 and listeria monocytogenesis so as to capture target bacteria from a detection sample; obtaining the target bacteria by magnetic field separation; treating by adopting propylmercuric bromide azide (PMA) to eliminate the interference of killed bacteria; and finally, extracting DNA (Deoxyribose Nucleic Acid) to carry out multiple PCR (Polymerase Chain Reaction) detection. Compared with a classical bacteria separation and identifying and serological typing method, the method disclosed by the invention is quick and accurate, can be used for only detecting viable bacteria and can be used for simultaneously detecting and identifying the three pathogens.

The invention discloses a bacteriophage capable of controlling Salmonella typhimurium; the bacteriophage is classified and named Salmonella typhimuriumphage Phi Sa-1, and is already collected in China Center for Type Culture Collection on 28 April, 2017, under CCTCC M2017217 in Wuhan University, China. The bacteriophage of the invention has specific killing activity for Salmonella typhimurium, and is capable of selectively killing specific pathogenic bacteria without killing beneficial bacteria or causing drug resistance. The bacteriophage also has excellent acid tolerance and heat resistance, may be prepared into feed additives, drinking water additives, feeds, drinking water, detergents or sanitizers and other various products, and is capable of killing Salmonella typhimurium under wide temperature range and pH range.

The invention discloses a method for detecting salmonella and staphylococcus aureus in food simultaneously. The method includes the following steps: 1), diluting a to-be-tested food sample according to GB4789 to obtain sample homogenate of 1:10, adding 50-65wt% of nitrilotriacetic acid solution into the sample homogenate according to the volume ratio of the sample homogenate to the nitrilotriacetic acid solution being (5-7):1, vibrating the mixture, adding phosphate buffer solution or 0.05% tween-20 contained phosphate buffer solution, performing centrifugal layering, and discarding an upper layer; 2), mixing a sample obtained after processing of the step 1) with immunomagnetic beads, vibrating the mixture, subjecting the mixture to standing and layering on a magnetic frame, discarding an upper layer, and washing a lower layer of the sample, wherein the immunomagnetic beads are obtained by modifying salmonella and staphylococcus aureus antibodies, which are labeled by magnetic bead coupled biotin, by streptavidin; 3), adding the phosphate buffer solution to the washed sample, vibrating the mixture, applying the mixture to an LB plate, and performing cultivation and observation. The method is simple and convenient to operate, accurate, efficient and low in cost, two target bacteria are enriched, and detection time can be shorten from 72h to 25h.

The invention discloses a salmonella enteritidis double knockout attenuated mutant and preparation as well as application thereof. The salmonella enteritidis double knockout attenuated mutant is obtained by knocking out crp gene and spiC gene of salmonella enteritidis C50041. The invention further discloses a preparation method as well as application of the salmonella enteritidis double knockout attenuated mutant. Further attenuation of the salmonella enteritidis attenuated mutant is realized. A foundation is laid for researching salmonella enteritidis attenuated live vaccines and live vector vaccines.

The invention relates to a primer combination for detecting infectious diarrhea pathogen. The primer combination comprises at least one combination of the following combinations: a rotavirus primer combination, an enteric adenovirus primer combination, a Norovirus primer combination, a salmonella primer combination, a Shigella primer combination and a campylobacter jejuni primer combination. The invention also relates to a kit containing the primer combination. The kit also comprises a primer-coating micro-fluidic chip, an isothermal amplification reaction liquid, an isothermal amplification enzyme solution, and a negative control. The invention also relates to a detection method by using the above primer combination. The detection method comprises steps as follows: coating of the primer combination, nucleic acid extraction of a sample to be detected, LAMP reaction and result interpretation. The kit provided by the invention can detect infectious diarrhea pathogen rapidly and accurately within one hour, and is also of great significance for rapid assisted guide treatment and drug use. The multi-index detection also can be used in regional epidemiological investigation and epidemic surveillance so as to study epidemic situation of infectious diarrhea in China.

The invention belongs to the technical field of biology and particularly relates to endolysin sourced from salmonella bacteriophage and an application thereof. The amino acid sequence of the endolysin sourced from the salmonella bacteriophage includes the total of the partial of the amino acid sequence represented as the SEQ ID No.1. Sequence structure analysis proves that the protease is a lyase which can lysing bacterial cell walls in the bacteriophage. A test proves that the protein formed by the amino acid sequence is good in bactericidal activity. The endolysin, when being expressed and purified by a suitable expression vector on market, can achieve a significant sterilization effect, so that the endolysin can be used for preparing a bacterial inhibitor of salmonella.

The invention provides Salmonella typimurium S496. The Salmonella typimurium S496 is characterized by lacking an rfbB gene and an rffG gene; a pagL gene is replaced by an arabinose-regulatory-sequence-containing rfbB gene with a changed SD sequence and a changed initiation codon, and the nucleotide sequence of the arabinose-regulatory-sequence-containing rfbB gene with the changed SD sequence and the changed initiation codon is shown as SEQ ID No.1; the classification of the attenuation strain is named as the Salmonella typimurium S496, the Salmonella typimurium S496 is collected in the China Center For Type Culture Collection (CCTCC), the collection number is CCTCC M2015562, and the collection time is September 21st, 2015. The invention further provides a method for constructing the Salmonella typimurium S496. Meanwhile, the invention provides an application of the strain or the attenuation strain constructed with the construction method in vaccine.

The invention discloses a Riemerella anatipestifer OmpA / MotB truncated recombinant protein, its antibody and a preparation method and an application thereof. The recombinant protein has immunoreactivity similar to natural OmpA / MotB protein, can bind to RA antibody specifically, will not generate cross reactions with Salmonella anatum positive serum, duck colibacilosis positive serum, duck swollen head septicemia virus positive serum, avian influenza virus (H5) positive serum, duck plague virus positive serum, duck hepatitis virus positive serum and the like, and can be used as a detection antigen for detecting whole bacteria antibody. As the recombinant protein is not whole bacteria, there is no risk of spreading bacteria when the recombinant protein is applied to detection. In addition, the preparation method of the recombinant protein is simple, is suitable for large-scale industrial production, can realize standardization of products and is beneficial to result comparison between different laboratories.

The invention discloses a preparation method and possible application of flagellin FliC from salmonella abortus equi to salmonella abortus equi immunity. Through cloning and connecting a flagellin FliC gene from the salmonella abortus equi to a prokaryotic expression vector pGEX-4T-2, a prokaryotic recombinant of a protein is obtained; the prokaryotic recombinant is transformed into engineering bacteria BL-21, and the protein is subjected to induced expression and purification; the protein has an amino acid sequence in a sequence table 1; the purified FliC recombinant protein has good immunogenicity after being used for the immunization of experimental animals, can overcome the defects of low antibody level and poor safety generated by inactivated vaccines and can be used for preparing subunit vaccines for preventing and treating equine paratyphoid infection and can be used as equine animal vaccine adjuvants, thereby having good development and application prospect.

The invention provides an antibacterial peptide and application thereof, and belongs to the technical field of biology. The antibacterial peptide is designed and obtained through a rational moleculardesign method on the basis of family analogues Pexiganan of the antibacterial peptide magainin derived from the epithelium of xenopus laevis. The antibacterial peptide is an antibacterial peptide Pexi-1 with an amino acid sequence as shown in SEQ ID NO. 1 or an antibacterial peptide Pexi-2 with an amino acid sequence as shown in SEQ ID NO. 2. The antibacterial peptide provided by the invention hasgood structural characteristics, can significantly improve the antibacterial effect on staphylococcus aureus, salmonella and aspergillus flavus, effectively reduces the hemolysis rate and cytotoxicity of chicken erythrocytes, and can be effectively used in animal-related production activities.

The invention discloses rare earth metal complexes using orbifloxacin as a ligand, a method for synthesizing the same and application thereof. In the tests of the inhibiting effect of the complexes on bacteria such as salmonella pullorum, swine streptococcosis and bovine streptococcus uberis, the results show that the complexes have quite high bacteriostatic activity, wherein all of Er(III), Sm(III) and Eu(III) complexes of the orbifloxacin have certain bacteriostatic activity on the swine streptococcosis which cannot be inhibited by orbifloxacin and doxycycline. Mouse acute toxicity tests show that the complexes have low in-vivo toxicity to mice and are highly tolerable to mice. The toxicity of various complexes meets the acute oral toxicity ungraded standards of European Community and the PRC food security assessment grade-two (actually non-toxic) standard, wherein the neodymium complex of the orbifloxacin has the lowest toxicity.

The invention discloses a chicken white diarrhea dyed condensation antigen, a preparation method and an application thereof and belongs to the technical field of veterinarian diagnosis. The antigen is prepared by following steps: performing recovery passage to chicken white diarrhea salmonella strains comprising SP7220, SP7701, SP8441 and SP9905 to obtain a seed bacterial liquid; inoculating the seed bacterial liquid on a solid culture medium for proliferation; inactivating the bacteria in formalin; regulating the concentration of the bacterial liquid through turbidimetry; and finally adding a crystal violet solution to dye the bacterial liquid, fully mixing uniformly the bacterial liquid and packaging the bacterial liquid to obtain the antigen. The preparation method is simple and reasonable, is scientific, is stable in production and is low in cost. The chicken white diarrhea condensation antigen production strain is good in antigenicity and is low in mutation rate. A prepared chicken white diarrhea dyed condensation antigen product is high in sensitivity, is strong in specificity, is quick in diagnosis and is clear in condensation image, and is an excellent dyed condensation antigen for detecting chicken white diarrhea.

The invention discloses a recombination attenuated salmonella typhimurium carrier vaccine of expression IBDV (Infectious Bursal Disease Virus) immunogenic gene, a preparation method and application thereof, in particular relates to oral attenuated salmonella typhimurium. The recombination attenuated salmonella typhimurium comprises a sequence disclosed by SEQ ID NO.1, and the recombination virus can express IBDV capsid protein VP2. The preparation method comprises the following steps of: inoculating the attenuated salmonella typhimurium in an LB substrate for culturing 18 hours; regulating bacterial concentration as 1010CFU / ml; and preparing the safe and effective oral attenuated salmonella typhimurium carrier vaccine for preventing IBD (Infectious Bursal Diseases).

The invention belongs to the technical field of medicine production research development, and relates to isolation and identification as well as application of volatile oil elements of Parasenecio firmus leaf. The isolation and identification of the volatile oil elements of the Parasenecio firmus leaf are realized by the following steps of: weighing dried Parasenecio firmus leaf, and extracting ultrasonically by a water steam distillation method to obtain the volatile oil; and analyzing the obtain volatile oil by utilizing a 6890 / 5973N gas chromatograph-mass spectrometer. The obtain volatile oil mainly comprises the following chemical elements in proportion: 18.98% of n-palmitic acid, 9.89% of 1-tridecylene, 8.05% of caryophyllene epoxide, 6.04% of (S)-undeca-cycloolefin, 3.84% of 9,12,15-octadecatrienoic acid, 2.97% of (-)-spathulenol, 2.83% of kaurene and 2.12% of caryophyllene. The volatile oil is used for preparing pharmaceutical preparation capable of restraining escherichia coli, streptococcus, Salmonella, Pasteurella and staphylococcus aureus.

The invention relates to a test paper box which is based on a bacterial fluorescent staining technique and is used for detecting dyed salmonella typhimurium via immunochromatography by utilizing a single antibody. The test paper box comprises a single antibody test strip and a fluorochrome box. The test paper box has the advantages that the salmonella typhimurium is dyed in advance by utilizing fluorochrome and then is sprayed on a T line of the test strip by utilizing a specific antibody of the salmonella typhimurium so as to capture the salmonella typhimurium in a mixed sample, so that the salmonella typhimurium in the sample can be rapidly and specifically detected.

The invention discloses a salmonella source tracing typing method based on a gMLST (genome multilocus sequence typing) technology. The method comprises the steps as follows: (1) isolation and dentification of salmonella; (2) DNA extraction and content detection; (3) whole-genome sequencing and gMLST typing; (4) MLST typing; (5) drug resistance gene analysis and virulence gene analysis; and (6) cladogram analysis. The method establishes a salmonella gMLST technology for important food-borne pathogens such as salmonella, staphylococcus aureus or vibrio parahaemolyticus to be applied to epidemiological source tracing, so that the method is promoted to source tracing of food-borne pathogens.

The invention discloses a preparation for producing a COVID-19 antibody after oral administration and a preparation method of the preparation. Attenuated salmonella typhimurium is used as a carrier todeliver DNA vaccine, the preparation effectively reaches intestinal tract and delivers DNA vaccine to intestinal endothelial cells with low mammalian toxicity infection through oral enteric-coated capsules, bacteria is phagocytosed through macrophages, and specific molecules COVID-19-S is expressed to be recognized by immune cells, so that the immune effect of specific antibodies in mammals (mice, humans and other mammals) neutralizing COVID-19-S protein is achieved.

The invention provides a detection test strip of salmonella typhimurium. The detection test strip comprises a sample pad, a cellulose nitrate membrane, absorbent paper and a bottom plate. A quantum dot fluorescent micro-sphere is marked with a monoclonal antibody of salmonella typhimurium, and the cellulose nitrate membrane is coated with a detection antibody and a quality control antibody. Basedon the test strip, the invention also provides a method for high-sensitivity and quick quantitative detection of salmonella typhimurium. The method fully utilizes super-strong and stable fluorescent signals of quantum dots, combines the advantages of easy marking and control of polymer sub-micro-spheres, constructs an immunochromatography technology of quantum dot fluorescent micro-spheres, and realizes sensitive and quick detection of salmonella typhimurium.

The invention discloses a primer, a probe and a chip for detecting and identifying a salmonella typhosa nucleic acid segment. The primer sequence comprises a primer pair sequence comprising an upstream primer sequence shown in SEQ ID NO.2 and a downstream primer sequence shown in SEQ ID NO.3. The probe sequence comprises 5'-Zc-P-3', wherein Zc at the 5' end is a sequence complementary to a probe Z fixed on a gene chip, and P at the 3' end is a specific sequence of a salmonella typhosa gene shown in SEQ ID NO.4. The gene chip comprises a probe for detecting and identifying the salmonella typhosa nucleic acid segment and a probe Z fixed on a solid-phase carrier. The primer, the probe and the chip provided by the invention can specifically and purposively detect the salmonella typhosa.

The invention discloses a novel chimeric cell-penetrating antibacterial peptide B6N2. The amino acid sequence is as shown in SEQ ID NO:1, the chimeric cell-penetrating antibacterial peptide is a chimeric polypeptide, and cell-penetrating peptides bLFcin6 with six amino acids are connected based on high-activity antibacterial peptide N2 to form the chimeric polypeptide. The antibacterial peptide B6N2 is simple in structure, low in toxicity and cost, safe and relatively easy to synthesize and can efficiently penetrate cells and kill intracellular salmonellae. The antibacterial peptide B6N2 is applied to the fields such as antibacterial drugs, food additives, cosmetics and feed additives and has wide application values and market prospect.

The invention relates to Salmonella typhi Ty21a comprising core-linked Shigella dysenteriae serotype 1 O-specific polysaccharide (O-Ps) and DNA encoding O antigen biosynthesis, said DNA selected from the group consisting of: a) the DNA sequence set out in any one of SEQ ID NOs: 1 and 2 and species homologs thereof; b) DNA encoding Shigella dysenteriae serotype 1 polypeptides encoded by any one of SEQ ID NOs: 1 and 2, and species homologs thereof; and c) DNA encoding a O antigen biosynthesis gene product that hybridizes under moderately stringent conditions to the DNA of (a) or (b); and related sequences, compositions of matter, vaccines, methods of using, and methods of making.

The invention discloses an oral recombinant attenuated salmonella typhimurium live vector vaccine as well as a preparation method and application thereof, particularly relating to an oral attenuated salmonella typhimurium. The recombinant attenuated salmonella typhimurium has a sequence shown in SEQ ID NO.1, and can express PCV-2 capsid protein ORF2. The attenuated salmonella typhimurium is inoculated in an LB culture medium for culturing for 18 hours; and the concentration of the attenuated salmonella typhimurium is regulated to 1010CFU / ml to prepare a safe and effective oral attenuated salmonella typhimurium live vector vaccine used for preventing porcine circovirus diseases.

The invention discloses an indirect ELISA kit for detecting a salmonella abortus equi antibody, and a preparation method and application of the indirect ELISA kit. The kit comprises an elisa plate coated with a salmonella abortus equi FljB protein, wherein the salmonella abortus equi FljB protein is coded by a sequence shown in SEQ ID NO.2. According to the indirect ELISA kit and method of the invention, sequence analysis is carried out, the specific sequence of the salmonella abortus equi is amplified by using P3 / P4, is cloned to a prokaryotic expression vector pET-28a; a recombinant expression plasmid pET28a-FljB is constructed, and is transferred into Rosetta (DE3); iPTG is used for inducible expression; the purified FljB protein is used as a coating antigen to establish the indirect ELISA method of the salmonella abortus equi antibody; and results show that the indirect ELISA method established by using the protein as the coating antigen has good specificity, sensitivity and repeatability, and can be used for clinical detection of the salmonella abortus equi antibody.

The invention discloses a method for rapidly detecting the concentration of salmonellae in water. The method comprises the following steps of: (1) obtaining a plurality of parts of salmonella solutions with different known concentrations; (2) rapidly detecting the concentration of the salmonellae by using an electrochemical chronoamperometry through a CHI660D electrochemical workstation; determining that a compliance relation between the salmonellae and boron-doped diamond film electrode responding current is a linear relation, wherein parameters of the CHI660D electrochemical workstation are that: a boron-doped diamond film electrode is used as a working electrode, the detection potential is 0.9-1.15V and an electrolytic tank is filled with a PBS (Phosphate Buffer Solution) with the pH (Potential of Hydrogen) being 7.2-7.6; and (3) carrying out sample introduction on a water body to be detected and recording a current value of responding electrophoresis of the boron-doped diamond film electrode; and detecting the salmonella concentration corresponding to the current value by utilizing an obtained responding current-salmonella concentration curve and combining the current value of the sample. The method disclosed by the invention is simple to operate; the detection time is only 1 hour; and the detection limit can reach 1000cfu / mL.

The invention discloses a PCR reaction tube coated with a specific antibody and used for detecting salmonella choleraesuis and an immune PCR detection kit. The PCR reaction tube is pre-coated with the monoclonal antibody which can specifically enrich salmonella choleraesuis in samples. The detection kit can fast, accurately and sensitively detect salmonella choleraesuis from food and other samples, and the sensitivity can reach 103-104 cfu / ml and is increased by 10-100 times than the sensitivity of a conventional PCR. An immunology method and a molecular biology method are organically combined into a whole, enrichment and detection of salmonella choleraesuis in the samples can be achieved in one PCR tube, the operation is simple, the cost is low, the detection is fast, and results are accurate.

The invention discloses a complex enrichment medium used for salmonella, listeria monocytogenes and staphylococcus aureus and a method for preparing the same. The complex enrichment medium comprises the following components in weight portion: 15 to 19 portions of tryptone, 2 to 4 portions of peptone, 2 to 3 portions of sodium dihydrogen phosphate, 2 to 3 portions of glucose, 0.5 to 1.5 portions of aesculin, 1, 000 portions of distilled water, 1 to 2.5 portions of sodium pyruvate, 3 to 10.0 portions of mannitol, 1.0 to 25.0 portions of sodium chloride, 1 to 2.5 portions of lithium chloride, 0.0001 to 0.0002 portion of potassium tellurite, and 0.005-0.015 portion of nalidixic acid, wherein the pH value is between 7.1 and 7.5. The complex enrichment medium can inhibit the growth of other pathogenic microorganisms while performing enrichment on three target pathogens, thus the complex enrichment medium can be directly applied to the isolation culture and the biological assay test of target bacteria, and can also be directly applied to a detection technology of a plurality of pathogens based on one detection platform for multiple PCR and the like to make a diagnostic report.

The invention discloses an application of a recombinant salmonella containing a nucleotide sequence shown in SEQ ID NO:1 in preparation of a medicine for preventing a porcine epizootic diarrhea disease and also discloses a vaccine which is used for preventing the porcine epizootic diarrhea disease and is prepared from the recombinant salmonella containing the nucleotide sequence shown in SEQ ID NO:1 and pharmaceutically acceptable auxiliary materials or a carrier. The vaccine for preventing the porcine epizootic diarrhea disease can produce a high-level antibody specific to porcine epizootic diarrhea viruses, has a good protective effect on the porcine epizootic diarrhea disease and has a good clinical application prospect.

The invention discloses a Riemerella anatipestifer OmpA / MotB signal peptide-removal recombinant protein, its antibody and a preparation method and an application thereof. The recombinant protein has immunoreactivity similar to natural OmpA / MotB protein, can bind to RA antibody specifically, will not generate cross reactions with Salmonella anatum positive serum, duck colibacilosis positive serum, duck swollen head septicemia virus positive serum, avian influenza virus (H5) positive serum, duck plague virus positive serum, duck hepatitis virus positive serum and the like, and can be used as a detection antigen for detecting whole bacteria antibody. As the recombinant protein is not whole bacteria, there is no risk of spreading bacteria when the recombinant protein is applied to detection. In addition, the preparation method of the recombinant protein is simple, is suitable for large-scale industrial production, can realize standardization of products and is beneficial to result comparison between different laboratories.

The invention relates to a swine cholera attenuated salmonella recombinant strain for expressing a haemophilus parasuis Omp 26 gene. Omp 26 is selected as an exogenous gene, a non-resistant plasmid pYA3493 is selected as a vector, and a swine cholera attenuated salmonella C500 asd gene deletion strain is adopted as a receptor strain to form an Asd+ balanced expression system. The asd gene can be stably inherited under the condition of no exogenous selection pressure, the problem of previous unstable inheritance of exogenous genes can be solved, the system has no marker of any antibiotic gene,a biological safety problem cannot be generated, and foundation is laid for developing the system into a safe, efficient and live vaccine vector; meanwhile, the outer membrane protein Omp 26 of haemophilus parasuis (HPS) can be well expressed and can also serve as special protein for researching clinical detection, epidemiological investigation and immune detection of the HPS. A basic idea is provided for research on bivalent vaccines of swine cholera and haemophilus parasuis.