32 results about "Cronobacter" patented technology

The invention discloses a nucleotide specific to a cronobacter O antigen and an application of the nucleotide. The nucleotide is at least one of nucleotides as shown in SEQ ID NO: 1-12 or at least one of complementary nucleotides. The specific nucleotide can be used for preparing a PCR (Polymerase Chain Reaction) kit and/or a gene chip for quickly detecting the cronobacters. The kit is simple and convenient, high in accuracy, good in sensitivity, excellent in operability, good in repeatability and low in detection cost if being applied to detecting clinical cronobacters or cronobacters in food.

The invention provides a tracing method of cronobacter spp. in infant formula milk powder. The method comprises the steps that by means of a multiple-locus sequence typing technology, molecular typing research is conducted on the cronobacter spp. of PIF raw materials, semi-finished products, finished products and processing environments thereof, population characteristics of the cronobacter spp. are revealed, a phylogenetic relationship of the cronobacter spp. is confirmed, phylogenetic analysis is conducted on the cronobacter spp. through bioinformatics, and therefore genetic characteristics of the cronobacter spp. in the above-mentioned environments are revealed systematically; on the basis of cronobacter spp. MLST typing, correlation analysis is conducted by taking ST of pathogenic bacteria and environmental sources as variables, key links which are most prone to pollute the cronobacter spp. in the PIF processing process and distribution rules are determined, and therefore the source of the cronobacter spp. in the infant formula milk powder is traced. The purpose of fundamentally monitoring the PIF production process and ensuring the PIF quality are achieved.

The invention relates to a detection medium and a detection method for Cronobacter in food. The detection method of the present invention utilizes the activity of alpha-glucosidase produced by Cronobacter to detect Cronobacter in liquid culture medium, and can identify Cronobacter while carrying out selective enrichment. The Cronobacter broth medium of the present invention may comprise 18-20 parts by weight of mixed peptone; 3-5 parts by weight of lactose; 2-3 parts by weight of K2HPO4; 2-3 parts by weight of KH2PO4; 20-34 parts by weight of sodium chloride 0.08-0.15 parts by weight of sodium lauryl sulfate; 0.009-0.011 parts by weight of vancomycin; 0.07-0.10 parts by weight of 4-methylumbelliferone-α-D-glucopyranoside; pH6.8 ± 0.2, the best Culture temperature, 36°C. The invention also discloses a reagent kit for the above detection method. The invention has the advantages of shortening the detection time and avoiding subsequent tedious identification operation steps, thereby improving the detection efficiency of Cronobacter.

The invention discloses a primer probe for RNA isothermal amplification to detect cronobacter, a kit and a detection method. The provided primer probe for detecting specificity, the detection kit containing the primer probe, and the detection method using the detection kit to carry out RNA isothermal amplification have the characteristics of high sensitivity, strong specificity, low pollution (amplification products RNA are easily degraded in nature), and rapidness; the requirements on instruments are low; the operation is simple; the primer probe and the kit are suitable for self detection offood enterprises and basic detection mechanisms and onsite detection, in particular, milk powder rapid detection, and the safety of milk powder is guaranteed.

The invention relates to a method for specific detection of Cronobacter spp. with silica magnetic microballoon and nested PCR technology based on a Cronobacter spp. 16SrDNA gene specific sequence, which is shown in the SEQID NO.1. Under specific condition, the property that silica magnetic microballoons can absorb nucleic acids is used for realizing rapid adsorption, enrichment and separation of pathogen nucleic acids in milk powder samples; two pairs of highly specific primers are designed based on Cronobacter spp. 16SrDNA gene, Cronobacter spp. specific gene is amplified by nested PCR technology, thereby realizing accurate, rapid and high sensitivity detection of Cronobacter spp. existed in the samples.

The invention provides application of a reagent for detecting flora in preparation of a reagent or a kit for prognosis prediction markers of esophageal squamous cell carcinoma patients, and belongs tothe technical field of prognosis detection of esophageal squamous cell carcinoma. In the present invention, the flora comprises one or multiple selected from Cronobacter in cancer tissues of patientswith esophageal squamous cell carcinoma, Family Halomonadaceae in para-carcinoma tissue of esophageal squamous cell carcinoma patients, and Carnobacterium in para-carcinoma tissue of esophageal squamous cell carcinoma patients. According to the invention, microorganism-related evidences are provided for prognosis prediction of esophageal squamous cell carcinoma, and a reference basis is providedfor prognosis prediction of esophageal squamous cell carcinoma patients.

The invention discloses a rapid detection method of enterobacter sakazakii in dairy products. The rapid detection method comprises the following steps: firstly, extracting dairy product samples and extracting RNA (ribonucleic acid) of more dairy product samples; adding 1-5 micrograms of the RNA in step 1 into a 0.5ml microcentrifuge tube and supplementing DEPCH2O to enable the total volume to reach 11 mu l; adding 10 microns Oligo (dt)12-18 microliters into the tube; then uniformly shaking and centrifuging; after heating at 70 DEG C for 10 min, immediately inserting the microcentrifuge tube instep 2 into an ice bath for 1 min. According to the rapid detection method of the enterobacter sakazakii in the dairy products, disclosed by the invention, mRNA (messenger ribonucleic acid) is used as a target gene for detection and can be detected through methods including PCR (polymerase chain reaction) and the like; the mRNA can be rapidly degraded after bacteria are dead, so that the proper mRNA is selected as a target gene and only the bacteria with activity can be detected, and false positive interference caused by dead bacteria is avoided; and meanwhile, a PCR primer is improved and designed and a real-time fluorescent RT-PCR technology is applied, so that combined detection of various bacteria can be realized and the time and the cost are saved.

The invention discloses a sensitization test strip for rapidly detecting cronobacter sakazakii and application of the sensitization test strip, and belongs to the technical field of detection of cronobacter sakazakii. The sensitization test strip comprises a sample pad, a gold standard pad, an NC film, an absorbent pad and a bottom plate, wherein the NC film is provided with a detection line and acontrol line, the control line is coated with a goat anti-mouse secondary antibody, and a detection line solution coating the detection line is obtained by incubating colloidal gold and an anti-cronobacter sakazakii capture antibody. The sensitization test strip has the characteristic of being capable of improving the protein binding quantity on the NC film, thereby enhancing the capture rate; the colloidal gold added at the detection line can improve aggregation coloration, thereby improving the detection effect; and the sensitivity is improved by 100 times compared with a traditional test strip, and the sensitization test strip has high specificity, short detection time, easy manufacture and a low cost, and is easy to carry and suitable for on-site detection of cronobacter sakazakii.

The invention discloses application of epsilon-polylysine or hydrochloride thereof in preparation of a medicine for inhibiting cronobacter spp or intervening a cronobacter spp biofilm. Experimental results show that epsilon-polylysine has the effects of inhibiting formation of the cronobacter spp and the biofilm thereof and removing the mature biofilm of the cronobacter spp. According to the invention, the bacteriostatic activity of the epsilon-polylysine to the cronobacter spp is determined by adopting a standard trace broth dilution method, the MIC value is 0.125-0.25 mg/mL, and the MBC value is 0.125-0.50 mg/mL; and by a crystal violet staining method and scanning electron microscope observation, the invention finds that the epsilon-polylysine with sub-inhibitory concentration has a remarkable inhibition effect on formation of the cronobacter spp biofilm, and the epsilon-polylysine with high concentration can destroy the mature biofilm. The invention provides experimental basis for clinical application of the epsilon-polylysine in the aspect of preparation of drugs for inhibiting the cronobacter spp or intervening the biofilm of the cronobacter spp, and the epsilon-polylysine has good application prospects and huge potential value.

The invention discloses two attenuated lipid A producing Cronobacter mutant strains and application thereof, and belongs to the technical field of genetic engineering. Through knocking out lpxM gene fragments of C.sakazakii BAA894 and ATCC 51329 bateria, the outer mold seepage performance of the strains is improved; the resistance on cationic anti-microbial peptides is reduced; the strains can bemore easily killed by the anti-microbial peptides; the copying survival capability of the C.sakazakii BAA894 and ATCC 51329 in mouse RAW264.7 cells and the stimulation effect on the HEK4 cells can bereduced; the infection level of the strains in macrophages is reduced; the cell toxicity is weakened; certain help is realized in the later development of vaccine adjuvants.

The invention discloses a primer combination and a kit for simultaneously detecting cronobacter and salmonella and application of the primer combination and the kit and belongs to the technical field of food safety. In order to solve the problems existing in detection on the salmonella and the cronobacter by a traditional microbiological method, the invention provides a dual test strip based on a LAMP reaction and a primer combination as shown in SEQ ID NO: 1-8, and the dual test strip comprises the following steps: extracting DNAs (deoxyribonucleic acids) from the salmonella and the cronobacter, and amplifying a large number of target products through the LAMP reaction; and directly interpreting a result by naked eyes through the dual test strip constructed on the basis of preparation of stable colloidal gold, a gold-labeled antibody, a redissolution solution and a diluent. The dual test strip obtained by the invention has the advantages of simplicity in manufacturing, low cost, sensitivity in detection and the like, and can be used for simultaneously detecting salmonella and cronobacter pollution in foods and food processing processes, especially pathogenic bacterium pollution in infant formula milk powder.

The invention belongs to the technical field of microbiological detection, and particularly discloses a primer probe combination, a kit and a method for detecting cronobacter based on an RAA technology. The primer probe combination comprises a primer pair and a probe. The primer pair comprises an upstream primer and a downstream primer, the sequence of the upstream primer is as shown in SEQ ID NO.1, the sequence of the downstream primer is as shown in SEQ ID NO.2, and the sequence of the probe is as shown in SEQ ID NO.5. When the kit and the method are used for detecting cronobacter, the specificity is good, the sensitivity is high, the accuracy is high, the operation is simple, the cost is low, laboratory detection and on-site rapid detection can be carried out, and the kit and the method have a good application prospect.

The invention discloses a ginkgo leaf flavone purified product and antibacterial application thereof. A purification method comprises a two-step purification method of NKA-9 macroporous resin and polyamide resin. The antibacterial application comprises the application of inhibiting staphylococcus aureus, escherichia coli and cronobacter malonate. The ginkgo leaf flavone purified product disclosed by the invention has a better antibacterial effect on staphylococcus aureus, escherichia coli and cronobacter malonate than that of a crude extract.

The invention relates to a cleaning agent with a cronobacter spp. biofilm inhibiting effect and a preparation method and application of the cleaning agent. The cleaning agent is prepared from amino oligosaccharides or derivatives thereof or alginate oligosaccharides, an anionic surfactant, a zwitterionic surfactant, a nonionic surfactant, glycerin and water. The cleaning agent has a great cronobacter spp. biofilm inhibiting effect and is especially capable of effectively inhibiting bacterial adhesion in an early stage of film formation. Compared with a common cleaning agent on the market, thecleaning agent containing the amino oligosaccharides or derivatives thereof or alginate oligosaccharides, the anionic surfactant, the zwitterionic surfactant and the nonionic surfactant has the advantage that the biofilm inhibiting effect is remarkably improved.

The invention discloses a paper-based detection device for detecting cronobacter and application of the paper-based detection device, belonging to the technical field of food safety. In order to solve the problems existing in processes of detecting cronobacter by using a traditional microbiological method and a molecular biological method, the invention provides the paper-based detection device, and the detection principle of the paper-based detection device is that the cronobacter can generate intracellular enzyme alpha-glucosidase after ultrasonication treatment, a specific enzyme-substrate reaction can be carried out on the paper-based detection device modified by the substrate X(alpha)lc, and then the cronobacter can be quickly and visually qualified and quantified on the paper-based detection device. The paper-based detection device disclosed by the invention has the characteristics of simple manufacturing method, low cost, high device sensitivity and the like, and can be used for detecting cronobacter pollution in food and food processing processes in real time, especially cronobacter pollution in production of infant formula milk powder.

The invention relates to a cronobacter thelingiensis type 5 lipopolysaccharide O-antigen oligosaccharide fragment and a preparation method and application thereof. The prepared oligosaccharide fragment is a cronobacter thelingiensis type 5 lipopolysaccharide O-antigen tetrasaccharide repeating unit and is prepared by taking various protected monosaccharide modules as raw materials and carrying out glycosylation reaction through a sequential glycosylation strategy, the C-1 site of a pyranosyl group at the reducing end of the oligosaccharide fragment is modified by an amino alkane chain, and can be conjugated with proteins, lipids, polypeptides and the like to prepare oligosaccharide conjugates, and then related immunological research is carried out. The preparation method disclosed by the invention is reasonable in design, strong in operability, simple in separation, high in product yield and good in purity. Immunological research is carried out by synthesizing polysaccharide repeated fragments with different reducing ends, and the method has important significance on research and development of corresponding cronobacter carbohydrate vaccines.

The invention relates to a method for detecting cronobacter in infant formula milk powder, and belongs to the technical field of detection of biological products. The innovation lies in improvement of enrichment and separation of cronobacter in infant formula milk powder. A liquid culture medium (dry damage repair factors comprise 0.4-0.8 g/L of mannitol and 2.5-5.0 g/L of trehalose, and selective enrichment factors comprise 80-150 g/L of sucrose, 4-8 g/L of vancomycin and 0.3-0.6 g/L of sodium deoxycholate) is adopted for efficient enrichment, and a solid culture medium (selective components comprise 0.5-1.0 g/L of sodium deoxycholate and 80-100 mg/L of 5-bromo-4-chloro-3-indole-D-alpha-glucoside) is adopted for selective separation. The method has the advantages of high enrichment efficiency, strong selectivity, convenience in operation, short time and the like, and overcomes the defects of long time, low enrichment effect, complicated operation and the like of the existing detection method. The method is suitable for detecting cronobacter in formula milk powder, shortens the detection time, improves the detection accuracy and efficiency, reduces the detection cost, and has important significance in guaranteeing the safety of infant formula milk powder.