21 results about "Yersinia enterocolitica" patented technology

The invention provides a primer combination of a multiple detection kit for food-borne pathogenic bacteria and the multiple detection kit. The primer combination comprises probes and primers for detecting nine food-borne pathogenic bacteria: vibrio cholerae general type, vibrio cholerae O1 type, vibrio cholerae O139 type, vibrio parahaemolyticus, salmonella, yersinia enterocolitica, campylobacter, listeria monocytogenes and enterobacter sakakaakii, and the sequence is shown as SEQ1-31; the probe and the primer pair are used for detecting nine characteristic genes, namely a shigella universal type: ipaH, a diarrheogenic Escherichia coli universal type: eltA, sta3, sta and aggR, and enterohemorrhagic Escherichia coli: stx1, stx2, escV and rfbE, and the sequences of the probe and the primer pair are shown as SEQ32-64. According to the invention, by combining a digital PCR multiple coding technology, the detection of 12 food-borne pathogenic bacteria can be realized, and the sensitivity, the specificity and the accuracy are relatively good.

The invention provides a bacteriostatic agent aiming at yersinia enterocolitica and application, the bacteriostatic agent comprises a compound solution of lipoic acid and protocatechualdehyde, and when the lipoic acid and the protocatechualdehyde act synergistically, the minimum inhibitory concentrations to the yersinia enterocolitica are 0.625 mg / mL and 0.3125 mg / mL respectively. The yersinia enterocolitica biofilm is inhibited and / or removed through the lipoic acid and protocatechuic aldehyde compound liquid, and the problem that drugs for treating the yersinia enterocolitica infection are prone to drug resistance is solved.

The invention discloses a lactobacillus jensenii strain and application thereof in fermentation of wall-broken ganoderma lucidum spore powder. The preservation number of the strain is CGMCC (China General Microbiological Culture Collection Center) No.31484. The strain is separated and screened from vaginal secretions of healthy women at the childbearing age, is sensitive to various common antibiotics, and has strong lactic acid production capacity; a fermentation supernatant of the bacillus subtilis shows relatively strong inhibition capability on escherichia coli, pseudomonas aeruginosa, staphylococcus aureus, salmonella typhimurium, shigella dysenteriae, yersinia enterocolitica and gardnerella vaginalis, and has relatively strong adhesion capability, growth inhibition capability and apoptosis promotion effect on human cervical cancer cells. When the strain is used for fermenting wall-broken ganoderma lucidum spore powder, the content of amino acid and bioactive substances such as peptides, arachidonic acids, steroids and polyketones can be greatly increased, the strain has an obvious inhibition effect on DSS-induced mouse colitis, and the inflammatory factor level is remarkably reduced. The strain not only has significant application value in biological agents, but also opens up a new way for combined application of lactic acid bacteria and ganoderma spores.

Provided is an internal reference gene for the real-time quantitative PCR (RT-qPCR) of Yersinia enterocolitica, including one of or a combination of some of glnS, nuoB, glmS, gyrB, dnaK, and thrS genes. Further provided is a primer set for amplifying the internal reference gene for the RT-qPCR of Yersinia enterocolitica. Given that stable housekeeping genes of Yersinia enterocolitica remain unclear, internal reference genes with relatively stable expression at different culture temperatures are selected by screening for Yersinia enterocolitica. Specific detection primers and real and reliable data provide a reference basis for selecting housekeeping genes of Yersinia enterocolitica; moreover, the use thereof alone or in combination, as an internal reference gene, improves the data accuracy, stability, and reliability, thereby solving the problem that usually only empirically selecting 16s rRNA as a single internal reference gene in conventional RT-qPCR results in unstable research results.

The invention discloses a method for preparing a sheep-derived positive serum reference of yersinia enterocolitica, obtained positive serum and application of the positive serum, and belongs to the field of veterinary medicine products. The invention aims to solve the technical problem of how to obtain the yersinia enterocolitica sheep-derived positive serum reference which is stable in titer and character and can be used for specific detection of brucella diagnostic products. Therefore, the invention provides a method for preparing the yersinia enterocolitica sheep-derived positive serum reference, which comprises the following steps: immunizing a tested sheep by using an immunizing antigen, and collecting serum of the immunized tested sheep to obtain the yersinia enterocolitica sheep-derived positive serum reference. The immune antigen is an inactivated yersinia enterocolitica obtained by inactivating the yersinia enterocolitica. The positive serum reference can be effectively applied to quality control evaluation work of brucellosis related diagnosis products, and plays a positive role in prevention and control of brucellosis.

The invention discloses a dual digital PCR (polymerase chain reaction) detection method for monkey campylobacter jejuni and monkey enterocolitis yersinia, and relates to the technical field of microbiological detection, the detection method comprises the following specific steps: multi-source heterogeneous data collection: collecting monkey feces and intestinal tissue samples, extracting genome DNA (deoxyribonucleic acid) by using a phenol-chloroform method, detecting the concentration and purity of DNA, and detecting the concentration and purity of the DNA; after reaching the standard, storing at-20 DEG C for later use; according to the method, high-sensitivity and specific detection on the two pathogenic bacteria is realized through a combination of well-designed primers and probes, according to the method, a gyrA gene conserved region of monkey campylobacter jejuni CJ and a POL gene conserved region of monkey yersinia enterocolitica YE are used as detection targets, and specific fragments of 249bp and 182bp are amplified respectively; the method effectively avoids interference of non-specific amplification, can simultaneously detect two pathogenic bacteria in a single reaction system through a dual digital PCR technology, accurately calculates target gene copy number concentration through a Poisson distribution formula, and improves detection accuracy and reliability.

Described herein are compositions, methods, and kits for detecting diarrhea causing pathogens from patient, food, or environmental samples. One embodiment described herein is primer pairs and probes for multiplex polymerase chain reaction (PCR) based assays for the detection of diarrhea causing pathogens, such as Vibrio cholerae, Vibrio parahaemolyticus, Vibrio vulnificus, Plesiomonas shigelloides, Yersinia enterocolitica, or Aeromonas hydrophila. Other embodiments include methods and kits for detecting diarrhea causing pathogens.

The invention provides a blood microorganism-based autism prediction model and application thereof, the prediction model takes blood microorganisms as markers, and the detection rate or relative abundance of the blood microorganisms is reduced; the risk that the blood sample suffers from autism is predicted by increasing the detection rate of blood microorganisms II or enhancing the relative abundance; the first blood microorganisms comprise ochrobactrum, pseudoalteromonas sp.3J6, rhodococcus equi, parabacteroides dieldrii, trichoderma harzianum, enterobacter hormaechei, wolbachia, yersinia enterocolitica, treponema pallidum and Elizabeth mosquito-repellent bacteria; the blood microorganisms II comprise Klebsiella compacticola and bdellovibrio bacteriovorus. According to the method, the model is constructed based on the blood metagenome data of 1946 standard autism four families, the advantages of blood microorganisms are brought into full play, disease signals are sensitively and accurately captured, the method can be applied to clinical autism prediction, and early risk assessment and layering of diseases are achieved.

The invention is suitable for the technical field of cold-chain food safety, and provides a low-temperature double-chamber ultrasonic atomization anti-psychrophile composition and an incremental iteration preparation method thereof. The composition is stored in a chamber A and a chamber B, after the compositions in the two chambers enter a Y-shaped mixer according to the volume ratio of 1: (0.8-1.2), the compositions are atomized into 40-60 [mu] m fog drops through an atomization net, the 24-hour sterilization rate of the fog drops to listeria monocytogenes and yersinia enterocolitica at the temperature of 0-10 DEG C is larger than or equal to 99%, and the composition is suitable for household refrigerator disinfection, cold chain storage preservation, transportation and other scenes. According to the preparation method, on the basis of an incremental active learning framework, sequence-structure-energy-mechanism multi-dimensional features are fused, rapid iteration of a formula is achieved through a light-weight gradient elevator (LightGBM), a graph neural network (GNN) algorithm and NSGA-II multi-objective optimization, full-link tracing is achieved in combination with a Git-DVC Hash chain, and FDAGMLP and ISO / IEC 24028 standards are met.

The present invention discloses a monoclonal antibody CJ116 against pathogenic Campylobacter and its application. In the monoclonal antibody CJ116, the amino acid sequence of the heavy chain variable region is as shown in SEQ ID NO.1, and the amino acid sequence of the light chain variable region is as shown in SEQ ID NO.5. This monoclonal antibody CJ116 has high affinity and high specificity, and the detection sensitivity for Campylobacter jejuni and Campylobacter coli can reach 10<supgt;2< / supgt> CFU / mL, and the detection sensitivity for Campylobacter fetus, Campylobacter upsaliensis and Campylobacter lari is 10<supgt;3< / supgt> CFU / mL, but there is no cross-reaction with other common intestinal pathogenic bacteria such as pathogenic Escherichia coli, Salmonella enteritidis, Yersinia enterocolitica, Shigella flexneri, Shigella boydii and Staphylococcus aureus.

Described herein are compositions, methods, and kits for detecting diarrhea causing pathogens from patient, food, or environmental samples. One embodiment described herein is primer pairs and probes for multiplex polymerase chain reaction (PCR) based assays for the detection of gastrointestinal pathogens, such as Adenovirus, Astrovirus, Rotavirus, Norovirus, Sapovirus, Campylobacter species, Clostridium difficile, Enteroaggregative E. coli (EAEC), Enteropathogenic E. coli (EPEC), Enterotoxigenic E. coli (ETEC), Shiga-like toxin-producing E. coli (STEC), Shigella / Enteroinvasive E. coli (EIEC), E. coli O157, Listeria monocytogenes, Plesiomonas shigelloides, Salmonella species, Yersinia enterocolitica, Vibrio species, Cryptosporidium species, Cyclospora cayetanensis, Entamoeba histolytica, Giardia intestinalis, or Giardia lamblia. Other embodiments include methods and kits for detecting gastrointestinal pathogens.

The invention discloses a monoclonal antibody CJ116 for resisting pathogenic campylobacter and an application of the monoclonal antibody CJ116. In the monoclonal antibody CJ116, the amino acid sequence of a heavy chain variable region is as shown in SEQ ID NO.1, and the amino acid sequence of a light chain variable region is as shown in SEQ ID NO.5. The CJ116 monoclonal antibody has high affinity and high specificity, the detection sensitivity of the CJ116 monoclonal antibody to campylobacter jejuni and campylobacter colons can reach 10 < 2 > CFU / mL, and the detection sensitivity of the CJ116 monoclonal antibody to campylobacter fetus, campylobacter upsackii and campylobacter seagull is 10 < 3 > CFU / mL. However, the enteropathogenic bacteria have no cross reaction with other common enteropathogenic bacteria such as pathogenic Escherichia coli, salmonella enteritidis, yersinia enterocolitica, Shigella flexneri, Shigella baumannii and Staphylococcus aureus.

This invention provides a composition, kit, and application for triple real-time fluorescence PCR detection of pathogenic Yersinia pestis based on TaqMan probes. Specific primers and TaqMan probes are designed targeting the caf1 gene of Yersinia pestis, the foxA gene of Yersinia enterocolitica, and the inv gene of Yersinia pseudotuberculosis, enabling simultaneous detection of these three pathogens in a single reaction system. The optimized annealing temperature is 54℃, and the final probe concentration is 0.3 μM. This method exhibits good linearity (R²>0.99), with detection limits of 5×10² copies / μL for I9 and C4, and 1×10¹ copies / μL for F4. Intra-assay and inter-assay coefficients of variation are mostly below 2%, and there is no cross-reactivity with non-target bacteria. It demonstrates good sensitivity, specificity, and repeatability, and can be used for rapid detection of pathogenic Yersinia pestis in food, environmental, clinical, and animal-derived samples.

The invention discloses a method for preparing a yersinia enterocolitica bovine-derived positive serum reference and the obtained bovine-derived positive serum reference, and belongs to the technical field of veterinary biological products. The invention aims to solve the technical problem of how to obtain the yersinia enterocolitica bovine-derived positive serum reference which is stable in titer and character and can be used for specific detection of brucella diagnostic products. Therefore, the invention provides a method for preparing the yersinia enterocolitica bovine-derived positive serum reference, which comprises the following steps: immunizing a tested bovine by using an immunizing antigen, collecting serum of the immunized tested bovine, and treating the serum to obtain the yersinia enterocolitica bovine-derived positive serum reference. The immune antigen is inactivated yersinia enterocolitica. The Yersinia enterocolitica bovine-derived positive serum reference material fills the blank of the Yersinia enterocolitica bovine-derived positive serum reference material for specific evaluation of a Brucella diagnosis product.

The present invention relates to the field of microbial technology, and in particular to Bacillus sabdariffa MLL-5 and its composite bacterial agent and application. The strain MLL-5 of the present invention is isolated by a research team from the rumen contents of healthy cattle. Testing shows that the strain has good β-1,4-glucanase production ability and has antibacterial effects on Escherichia coli, Yersinia enterocolitica, Shigella flexneri and Salmonella typhimurium. The Bacillus sabdariffa MLL-5 has good fermentation ability for cassava residue, can effectively reduce the cyanide content of cassava residue fermented feed and increase the protein content of cassava residue fermented feed, and is a good biological fermentation agent for cassava residue. After feeding Jersey cattle with cassava residue fermented feed prepared by using the strain and the composite bacterial agent, it was found that adding 10%-15% of fermented cassava residue to the diet did not have an adverse effect on the growth performance and health of the cattle, and significantly reduced the number of diarrhea episodes. The Bacillus sabdariffa MLL-5 is a safe fermented feed.

The invention relates to the technical field of microorganisms, in particular to bacillus safensis MLL-5 and a complex microbial inoculant and application thereof.The strain MLL-5 is separated from rumen contents of healthy cattle by a research group, and it is detected that the strain has the good beta-1, 4-glucanase producing capacity and can be used for producing beta-1, 4-glucanase. The antibacterial effect is achieved on escherichia coli, yersinia enterocolitica, shigella flexneri and salmonella typhimurium; the bacillus safensis MLL-5 has relatively good fermentation capability on the cassava residues, can effectively reduce the cyanide content of the cassava residue fermented feed and improve the protein content of the cassava residue fermented feed, is a good biological fermentation agent aiming at the cassava residues, and has good application prospects. After Jersey cattle are fed with manioc waste fermented feed prepared from the strain and the complex microbial inoculant, it is found that addition of 10%-15% of fermented manioc waste into daily ration does not have adverse effects on the growth performance and health conditions of the cattle, the number of diarrhea times is obviously reduced, and the manioc waste fermented feed is safe.

This document describes compositions, methods, and kits for detecting diarrheal pathogens from patient, food, or environmental samples. One embodiment described herein is primer pairs and probes for a multiplex polymerase chain reaction (PCR)-based assay for detecting gastrointestinal pathogens such as adenovirus, astrovirus, rotavirus, norovirus, zaruzin virus, Campylobacter species, Clostridium difficile, enteroaggregative Escherichia coli (EAEC), enteropathogenic Escherichia coli (EPEC), enterotoxin-producing Escherichia coli (ETEC), Shigella-like toxin-producing Escherichia coli (STEC), Shigella spp. / enteroinvasive Escherichia coli (EIEC), Escherichia coli O157, Listeria monocytogenes, Shigella-like bacteria, Salmonella spp., Yersinia enterocolitica, Vibrio species, Cryptosporidium species, Cyclospora kayanoides, Entamoeba histolytica, Giardia lamblia, or Giardia lamblia. Other embodiments include methods and kits for detecting gastrointestinal pathogens.

The present invention provides a kit for joint detection of eight foodborne pathogenic bacteria based on the multiplex probe melting curve method, belonging to the technical field of microbial detection. The primer-probe Mix in the kit provided by the present invention includes primers shown in SEQ ID NO.1-16, probes shown in SEQ ID NO.17-24, and substrates shown in SEQ ID NO.25-32. The present invention uses 2 fluorescence channels and differentiates them through different Tm temperature peaks of substrate melting peaks in a single fluorescence channel. The present invention realizes the full detection of characteristic genes in 8 foodborne pathogenic bacteria (Salmonella, Staphylococcus aureus, Escherichia coli O157, Listeria monocytogenes, Vibrio, Yersinia enterocolitica, Cronobacter sakazakii, Shigella) in a single-tube fluorescence quantitative PCR reaction, and has the advantages of simple operation, good specificity, high sensitivity and low cost.

The application belongs to the technical field of microorganism detection, and specifically discloses a preparation and application of immunomagnetic beads for enriching Yersinia enterocolitica, wherein a recombinant protein Omp370-3 with an amino acid sequence shown as SEQ ID NO. 2 is used as an antigen to immunize animals to obtain Omp370-3 polyclonal antibodies; the Omp370-3 polyclonal antibodies are coupled with carboxyl magnetic beads to obtain immunomagnetic beads. The immunomagnetic beads have high efficient enrichment capacity for Yersinia enterocolitica, and the enrichment efficiency can reach 83%. The bacteria after enrichment can be directly applied to subsequent RAA detection, so that the detection time can be shortened, the detection sensitivity can be improved, and new technical support is provided for the detection of foodborne pathogenic bacteria in food (such as pork).

The application provides an autism prediction model based on blood microorganisms and application thereof. The prediction model takes blood microorganisms as markers, and predicts the risk of autism of a blood sample through the decrease of a blood microorganism one detection rate or relative abundance, or the increase of a blood microorganism two detection rate or relative abundance. The blood microorganism one comprises Xanthobacteraceae bacterium, Pseudomonas sp.3J6, Rhodococcus equi, Parabacteroides distasonis, Xylella fastidiosa, Enterorhabdus hofmannii, Wolbachia, Yersinia enterocolitica, Borrelia parkeri and Elizabethkingia sp. The blood microorganism two comprises Klebsiella michiganensis and Bdellovibrio sp. The application constructs a model based on blood metagenomic data of 1946 standard four-generation families of autism, fully develops the advantages of blood microorganisms, sensitively and accurately realizes the capture of disease signals, and can be applied to the prediction of autism in the clinic, and realizes the early risk assessment and stratification of diseases.

The invention relates to the technical field of microbiological detection, in particular to a method for enriching O: 3 type enterocolitis yersinia by Gp17 magnetic beads, which comprises the following steps: step 1, expressing and purifying Gp17 protein through a prokaryotic expression system; step 2, coupling the purified Gp17 protein in the step 1 with carboxyl modified magnetic nanoparticles to prepare Gp17 functionalized magnetic beads; step 3, mixing the Gp17 functionalized magnetic beads prepared in the step 2 with a sample to be detected, and enriching O: 3 type enterocolitis yersinia through magnetic field separation; 4, the enriched O: 3 type enterocolitis yersinia is cultured or subjected to molecular biology detection.The method for enriching the O: 3 type enterocolitis yersinia through the Gp17 magnetic beads has the advantages that compared with existing immunomagnetic beads and Gp17 functional magnetic beads, the specificity is better, enrichment of specific serotypes can be achieved, and the method is suitable for large-scale popularization and application. And the Gp17 protein can be massively produced in vitro by utilizing a prokaryotic expression technology and is easy to obtain, the cost is reduced, and the difference between batches is small.