88 results about "Yersinia enterocolitica" patented technology

The present invention demonstrated the potential use of Lactobacillus johnsonii D115 as a probiotic, as a prophylactic agent or as a surface treatment of materials against human and animal pathogens such as Brachyspira pilosicoli, Brachyspira hyodysenteriae, Shigella sonnei, Vibrio cholera, Vibrio parahaemolyticus, Campylobacter jejuni, Streptococcus pneumoniae, Enterococcus faecalis, Enterococcus faecium, Clostridium perfringens, Yersinia enterocolitica, Escherichia coli, Klebbsiella pneumoniae, Staphylococcus aureus, Salmonella spp., Bacillus cereus, Aspergillus niger and Fusarium chlamydosporum. The proteineous antimicrobial compound was partially characterized and found to be heat tolerant up to 121° C. for 15 min, and acid tolerant up to pH1 for 30 min at 40° C. The compound is also stable to enzymatic digestion, being able to retain more than 60% antimicrobial activity when treated with pepsin and trypsin.

The invention relates to a pathogenic microorganism DNA detection chip and the chip comprises a carrier and a nucleic acid probe on the carrier. The nucleic acid probe is provided with a target detection probe used for detecting target gene of pathogenic microorganism. The pathogenic microorganism comprises but not limits to vibrio cholerae, pathogenic escherichia coli, campylobacter jejuni, Yersinia enterocolitica, parahemolytic vibrio, salmonella, and shigella and Listeria monocytogenes. The invention also relates to a preparation method of the pathogenic microorganism DNA detection chip and provides applications of the pathogenic microorganism DNA detection chip in detection of pathogenic microorganism. Also a pathogenic microorganism DNA detection chip kit is provided.

The invention discloses a fourteen-food-borne pathogenic bacterium multiplex PCR detection primer set and a kit. The detection primer set is composed of primer pairs for detecting salmonella, Shigella, Vibrio parahemolyticus, campylobacter jejuni, campylobacter coli, staphylococcus aureus, bacillus cereus, Listeria monocytogenes, yersinia enterocolitica, enterobacter sakazakii, Escherichia coli, vibrio cholerae, Escherichia coli O157, aeromonas hydrophila and internal positive control. A multiplex PCR detection method based on an ordinary PCR platform is built, multiplex PCR reactions are carried out on genomic DNA, extracted from a sample to be tested, of bacteria in the same reaction system through the primer pairs acquired through analysis and design, and whether the food-borne pathogenic bacteria are contained in the sample or not is judged through the electrophoretic analysis of reaction products.

The present invention belongs to the field of microbe detecting technology. The oligonucleotide probe for detecting common intestinal tract pathogenic bacteria is designed on 16S rRNA and 23S rRNA of bacteria, ipaH of dysentery bacillus giant plasmid, VipR of Salmonella typhi and other gene sequence, has length of 25-50 bp, and relatively high sensitivity and specificity. The oligonucleotide probe is suitable for detection based on nucleic acid hybridization principle, especially detection based on gene chip principle. Under certain use condition, it can detect Listeria, parahemolutic vibrio, Campylobacter, etc. It may be used in many aspects, such as disease diagnosis, environment detection, food poisoning detection, etc.

The present invention relates to a safe non-virulent Yersinia enterocolitica mutant strain for delivering heterologous proteins in target cells carrying mutations in at least one of the effector genes yopH, yopO, yopP, yopE, yopM, yopT genes and at least one additional mutation in the invasin genes chosen from yadA and/or inv. The present invention also relates to a safe non-virulent Yersinia enterocolitica mutant strain for delivering heterologous proteins in target cells according to claim 1 carrying mutations in all effector genes yopH, yopO, yopP, yopE, yopM, yopT genes and at least one additional mutation in the invasin genes chosen from yadA and/or inv. The present invention also relates to an expression vector for delivering a heterologous protein into a target cell using a Yersinia enterocolitica mutant strain according to any of the claims 1 to 4, which comprises in the 5' to 3' direction :(a) a promoter of a Yersinia virulon gene, (b) a first DNA sequence encoding a delivery signal from a Yersinia effector protein, operably linked to said promoter; and, (c) a second DNA sequence coding for said heterologous protein, fused in frame to the 3' end of said first DNA sequence. The present invention further relates to methods and compositions comprising (the use of) the afore-mentioned mutant strains and expression vectors.

An immobilized microbial consortium is formulated which comprises of a synergistic mixture of all the bacterial strains of Aeromonas hydrophila, Pseudomonas fluorescens, Pseudomonas aeruginosa, Bacillus circulans, Yersinia enterocolitica, Enterobacter cloaca and Bacillus brevis. The formulated microbial consortium is immobilized on a non-biodegradable and economically cheaper support. The said immobilized microbial consortium is used for the biodegradation of synthetic phenol as well as phenol present in petroleum refinery effluent. The results of biodegradation obtained with the microbial consortium immobilized on coconut fiber are compared with those obtained with microbial consortium immobilized on well known support. The coconut fiber used for immobilization proved to be a better support than a well known support such as calcium alginate.

The invention provides a method for detecting fluorescent quantitative PCR of 12 pathogenic bacteria in real time at the same time, including the steps: collecting specific pathogenic gene or toxin gene of the target pathogen and using it as a target gene to design primers and probes so as to make the reaction conditions consistent; extracting a genome template of a sample to be detected; adding the template respectively into tubules equipped with different specific upstream and downstream primers and probes, and then adding the corresponding fluorescent quantitative PCR reagents; under the same cycle of fluorescent quantitative PCR, the corresponding primers and probes are used to detect the samples simultaneously, quickly and quantitatively in their respective reaction tubes. Easier, Quick and efficient, Twelve common pathogenic bacteria (Escherichia coli O157: H7, Listeria monocytogenes, Salmonella, Vibrio parahaemolyticus, Streptococcus betae, Yersinia enterocolitica, Streptococcusfaecalis, Shigella, Proteus mirabilis, Vibrio fluvialis, Campylobacter jejuni, Staphylococcus aureus) can be detected simultaneously in drinking water and food economically.

The invention provides a genetic chip and a detection kit related to major pathogenic microorganisms causing infectious diarrhea of human beings, which are mainly specific to 13 types of bacteria including lapactic bacillus coli/shigella, salmonella, comma bacillus, vibrio parahemolyticus, aeromona hydrophila, plesiomonas, virio hollisae, vibrio fluvialis, vibrio mimicus, Wauter's strains, vibriodamsela and vibrio furnissii. The genetic chip comprises a solid-phase carrier and an oligonucleotide probe fixed on the solid-phase carrier, wherein the oligonucleotide probe comprises DNA (Deoxyribonucleic Acid) fragments selected from genes including a gyrB gene, an ITS gene, a dnaJ gene, a toxR gene and the like with remarkable biological evolution advantages or complementary DNA fragments. By adopting the genetic chip and the detection kit provided by the invention, major pathogenic microorganisms causing infectious diarrhea of human beings can be detected. The genetic chip and the detection kit have the characteristics of easiness and convenience for operation, high flux, high accuracy, high repeatability and the like, and can be applied to clinical detection of inspection departments in hospitals.

The invention discloses a method by using composite fluorescence PCR technique to detect food-borne pathogenic enterobacter and pertains to bacteria detection technical field. The main technic proposal is to design a primer group sequence. The pathogenic enterobacter is common pathogenic bacteria in food and imposes a serious thread to human health. The quick and accurate detection of pathogenic enterobacter in food is a main premise condition for effective prevention and control of pathogen bacteria infection. The food-borne pathogenic enterobacter required detection mainly comprises shigella, salmonella, Yersinia enterocolitica, hemorrhagic scherichia coli and scherichia coli O157: H7. The invention overcomes technical shortage in the prior art aiming at the object bacteria and provides the quick and low-cost detection method by using composite fluorescence PCR technique to detect shigella, salmonella, Yersinia enterocolitica, hemorrhagic scherichia coli and scherichia coli O157: H7. The method can use two-diode PCR reaction and primarily screen shigella, salmonella, Yersinia enterocolitica, hemorrhagic scherichia coli and scherichia coli O157: H7.

The present invention relates to novel primers of SEQ ID Nos. 1-4 useful for detecting poisoning in food articles wherein primers of SEQ ID Nos. 1 and 2 are directed against enterotoxin A gene (ent A) of bacteria Staphylococcus aureus and primers of SEQ ID Nos. 3 and 4 are directed against heat stable enterotoxin gene (yst) of bacteria yersinia enterocolitica, and a highly sensitive method of detecting said food poisoning bacterial species using said primers.

The invention discloses universal primers and a probe for on-site rapid detection of Brucella and a kit. The forward primer sequence is shown as SEQIDNo.1, the reverse primer sequence is shown as SEQIDNo.2, and the probe sequence is shown as SEQIDNo.3. The universal primers, the probe and the kit provided by the invention for detection of Brucella have high sensitivity and strong specificity, minimumly can detect 6.0*10<0>cfu Brucella, and have no cross reaction with Escherichia coli O157:H7, yersinia enterocolitica O:9, Salmonella, Staphylococcus aureus and other bacteria. The universal primers, the probe and the kit provided by the invention not only can be used for detection of Brucella strains, but also can be used for detection of clinical samples, which mainly include blood, milk samples, aerosol samples, and tissue samples, etc. The kit provided by the invention is convenient to use, has no need for special equipment, can carry out sensitive, specific and rapid detection of Brucella on the crude split product of a to-be-detected sample in 25min just at 38DEG C, and is suitable for field or grassroots brucellosis quarantine work.

The invention relates to a fluorescence PCR (polymerase chain reaction) kit for detecting yersinia enterocolitica, reagents in the kit comprise a front primer, a rear primer, a TaqMan probe, a fluorescence PCR premixed solution, an ROX (roxithromycin) solution, pure water and a positive standard solution, wherein the front primer, the rear primer and the TaqMan probe are DNA (deoxyribonucleic acid) fragments of 20-21 bases, which are synthesized according to a specific and conserved gene nucleotide sequence of the yersinia enterocolitica and can be specifically combined with the yersinia enterocolitica. The invention further provides a method for detecting the yersinia enterocolitica, and the method comprises the following steps: performing pretreatment on samples; using the reagents of the kit for detection; and analyzing the detection result. The kit is simple, fast, sensitive, accurate and suitable for detecting a large number of samples.

The invention discloses a detection kit of 16 food born pathogenic bacteria. The detection kit comprises a PCR amplification reagent and reference reagents; the PCR amplification reagent is composed of PCR buffer solution A, enzyme system B, and a primer probe mixed solution of the 16 food-borne pathogenic bacteria; the reference reagents are mainly composed of a negative control and a positive control; the PCR buffer solution A contains 5*Buffer, 25mM dN(U)TPs; the enzyme system B is mainly composed of HotStart Hi Taq DNA polymerase and UNG anti-pollution enzyme; the primer probe mixed solution of the 16 food-borne pathogenic bacteria contains a mixed solution of detection primer probes of salmonella, Shigella, enteropathogenic Escherichia coli, enterotoxigenic E. coli, enteroinvasive escherichia coli, adherent escherichia coli, Escherichia coli O157, Staphylococcus aureus, listeria monocytogenes, Bacillus Cereus, campylobacter jejuni, vibrio parahaemolyticus, Vibrio cholerae group O1, Vibrio cholerae group O139, Yersinia enterocolitica, and enterobacter sakazakii. The detection kit is high in detection sensitivity, is simple, and is convenient.

Provided are novel primers directed against enterotoxin A gene (ent A) of bacteria Staphylococcus aureus and primers directed against heat stable enterotoxin gene (yst) of bacteria Yersinia enterocolitica, for detecting poisoning in food articles. Also provided is a highly sensitive method for detecting bacterial food poisoning using the primers.

The invention provides a genetic marker and a method for detection of yersinia enterocolitica, in particular a nucleotide sequence for detection of the ail gene of the yersinia enterocolitica, oligonucleotide primers and a probe. The oligonucleotide primers and the probe are designed based on the nucleotide sequence. The invention also provides a method for rapid and specific detection of the yersinia enterocolitica with the primers. Moreover, the invention can detect the yersinia enterocolitica conveniently, rapidly, accurately, qualitatively and quantitatively.

The invention discloses a method for detecting food origin enterocolitis Yersinia by using loop-mediated constant temperature PCR technology, belonging to the food sanitation field. The technical proposal is as follows: enterocolitis Yersinia 16S-23S regions are used as target detection sequences, and four specific primers containing the whole region sequences are designed. The invention comprises extraction of four specific primers of enterocolitis Yersinia 16S-23S regions and food origin enterocolitis Yersinia, detection method for loop-mediated constant temperature PCR amplification and interpretation of results. The method fills the gap of a method for detecting food origin enterocolitis Yersinia by using loop-mediated constant temperature PCR amplification technology, and is used for checking enterocolitis Yersinia in import and export foods. The method has the characteristics of low cost, high efficiency, simple operation and convenience.

Immobilized cell beads incorporating formulated microbial consortium comprising a synergistic mixture of the following bacterial strains namely, Enterobacter sakazaki, Pseudomonas aeruginosa and Aeromonas sobria selected from the following isolated bacterial strains namely, Yersinia enterocolitica, Aeromonas sobria, Klebsiella pneumoniae, Serratia liquefaciens, Enterobacter sakazaki, Citrobacter amalonaticus, Pseudomonas fluorescens, Pseudomonas aeruginosa, Enterobacter cloaca, Acinetobacter calcoaceticus are prepared, the formulated microbial consortium is immobilized in an appropriate immobilizing agent resulting in the formation of beads and the beads are used for instant BOD estimation using an electronic device and the formulated cell beads are reusable and are capable of assimilating most of the organic matter present in varied industrial effluents.

The invention relates to a diarrhea pathogenic bacteria multi-gene detection system and its kit and application. The detection system has 15 pairs of primers, wherein 13 pairs of diarrhea pathogens primers, one pair of human genome beta-actin primers and one pair of system quality controlled beta-actin primers are included. Diarrhea pathogens refer to campylobacter jejuni, shigella, clostridium difficile, salmonella enteritidis, salmonella typhimurium, enterotoxigenic escherichia coli, escherichia coli O157, vibriones, yersinia enterocolitica and the like. The diarrhea pathogenic bacteria multi-gene detection system and its kit do not require routine culture and other steps, synchronous detection and analysis of the various diarrhea pathogens can be directly conducted on a stool sample in the same reaction system, the shortcomings that a routine detection method is low in flux and low in detection rate and takes much time are made up for, a comprehensive, accurate and low-cost pathogen diagnosis is provided for clinical use for the first time, and an important reference is provided for individualized medication and accurate medical treatment.

The invention relates to a RCA (rolling circle amplification) rapid detection primer and kit for Yersinia enterocolitica, wherein the sequence of a primer combined with the specificity of a probe is SEQ ID NO: 1; the sequence of a primer combined with a fragment amplified through taking the probe as a template is SEQ ID NO: 2; and the sequence of the probe is SEQ ID NO: 3. The kit provided by the invention is characterized in that a padlock probe is designed according to the conserved gene sequence of a to-be-detected bacterial strain so as to ensure the specificity of a detection method. According to the invention, an improved rolling circle amplification (RCA) technique is adopted, and the technique is strong in specificity, and has higher sensitivity than that of a PCR (polymerase chain reaction) detection method; meanwhile, the technique is simple, rapid, safe, pollution-free, and especially applicable to food detection mechanisms.

The invention discloses a preparation method of a quick nucleic acid test strip detection kit for food-borne pathogenic microorganism-yersinia enterocolitica and an application thereof, and also discloses a design method of a primer for specific nucleic acid PCR (polymerase chain reaction) amplification for yersinia enterocolitica, and a method for detecting a PCR amplification product with the nucleic acid test strip for nucleic acid lateral flow and an application. The method disclosed by the invention can be used for detecting the amplification product of the target nucleotide sequence of pathogenic microorganisms from other sources by use of the specific nucleotide sequence amplification product, wherein the specificity is guaranteed by the amplification primer.

The invention discloses a common primer-mediated PCR kit for detecting 11 food-borne pathogenic bacteria and an application of the common primer-mediated PCR kit. A common primer-mediated multiplex PCR technology is utilized; specific genes of 11 common food-borne pathogenic bacteria (salmonella, listeria monocytogenes, shigella flexneri, enterohemorrhagic escherichia coli O157, vibrio parahaemolyticus, staphylococcus aureus, vibrio cholerae, clostridium botulinum A, bacillus cereus, clostridium perfringens and yersinia enterocolitica) are used as detection markers; an efficient, high-specificity and high-flux multiplex PCR detection system is established, and a nucleic acid detection kit suitable for detecting the common food-borne pathogenic bacteria is researched and developed. The sensitivity of the common primers reaches 10<3> copies/mu L, a UP-M-PCR system for simultaneously detecting the 11 food-borne pathogenic bacteria is constructed, single-template and two-template experiments prove that the UP-M-PCR system has good specificity, and the sensitivity of a bacterial genome DNA detection system is as high as 0.01 ng/mu L.

The invention discloses a method, a primer group and a kit for quickly detecting yersinia enterocolitica in a constant-temperature manner. The method comprises the following steps of extracting DNA (Deoxyribonucleic Acid) of a genome from a to-be-detected sample; using the DNA of the genome as a template, using a primer group capable of amplifying the specific sequence of the yersinia enterocolitica as primers, and carrying out constant-temperature amplification reaction under an enzyme reaction system; through judging whether a reaction result is positive or not, determining whether the yersinia enterocolitica exists in the to-be-detected sample or not. The detection method provided by the invention has high sensitivity and high specificity, is short in detection time, is simple in result decision, is convenient and quick to operate, is low in cost, and has a wide application prospect.

The invention discloses a detection kit for Yersinia rohdei causing enterocolitis. The detection kit comprises primer pairs shown as SEQ ID NO: (1 to 2), SEQ ID NO: (3 to 4) and SEQ ID NO: (5 to 6). The invention further provides a detection method of Yersinia rohdei, the three primer pairs, and application of the primer pairs. The detection kit and the detection method has the advantages that Yersinia rohdei can be detected accurately and effectively; the specificity and the sensitivity are high; the time consumption is low; the detection is fast; fast detection and forecast of water sources and foods can be realized; diseases can be prevented; the economic benefit is improved.