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Rapid PCR detection method and kit for klebsiella pneumoniae, acinetobacter baumannii and pseudomonas aeruginosa

Resumen de: CN121160897A

The invention discloses a rapid PCR (polymerase chain reaction) detection method and kit for klebsiella pneumoniae, acinetobacter baumannii and pseudomonas aeruginosa, and belongs to the technical field of microbiological detection. The kit comprises a primer probe combination which is specifically designed for an rpsD gene of klebsiella pneumoniae, an acinetobacter baumannii recA gene and a pseudomonas aeruginosa ecfx gene, and the combined probe is subjected to Spacer linker blocking modification, so that the detection specificity of a system is remarkably improved. The method is simple, convenient and rapid to operate, the detection sensitivity of the klebsiella pneumoniae, the acinetobacter baumannii and the pseudomonas aeruginosa can reach 0.5 copies/mu L, the specificity is good, and an effective tool is provided for early and rapid diagnosis of lower respiratory tract infection caused by the klebsiella pneumoniae, the acinetobacter baumannii and the pseudomonas aeruginosa clinically.

Multiplex fluorescent quantitative PCR (polymerase chain reaction) detection kit for prawn culture pathogenic bacteria

Resumen de: CN121160893A

The invention discloses a multiplex fluorescent quantitative PCR (polymerase chain reaction) detection kit for prawn culture pathogenic bacteria, which is characterized in that the kit contains primers and probes for detecting the prawn culture pathogenic bacteria, wherein the primers and the probes are respectively used for detecting TDH-related hemolsin genes of vibrio parahaemolyticus, ORF107 genes of white spot syndrome viruses, small subunit ribosomal genes of shrimp enterocytozoon hepatopenaei, 37 kDa coat protein genes of infectious subcutaneous and hematopoietic necrosis viruses, DNA (deoxyribonucleic acid) primer genes of full-eye iridovirus 1 and PirA pi genes of pathogens of bacterial prawns acute hepatopancreas necrosis; the method has the advantage that the detection of eight prawn culture pathogens can be realized at the same time. The method comprises the following steps of: obtaining a tcdB gene of a high-pathogenicity vibrio and a hemolsin gene of a vibrio harveyi;

PSEUDOMONAS AERUGINOSA INA PROTEIN AND USE THEREOF

Resumen de: WO2025256021A1

A Pseudomonas aeruginosa InA protein and a use thereof, relating to the technical field of biological detection. The InA protein is InAS2, InAAP41, or InAS8, the amino acid sequences of which are respectively as shown in SEQ ID NOs: 1-3; and nucleotide sequences encoding InAS2, InAAP41, and InAS8 are as shown in SEQ ID NOs: 4-6. Compared with the prior art, the InA protein can improve the solubility of a target protein. In addition, there is also an ultra-high affinity between the InA protein and a Pseudomonas aeruginosa immune protein, and on this basis, a reversible and regenerable biosensing chip is developed. The chip can capture a fusion protein carrying the InA protein and can be used for detecting the affinity between the fusion protein carrying an InA tag and a candidate drug, thereby achieving drug screening. Moreover, the protein-drug affinity kinetics detected by the chip can also be used in drug pharmacology research.

PRIMER AND PROBE FOR DETECTING SALMONELLA PULLORUM, AND KIT

Resumen de: WO2025255902A1

A primer and probe for detecting Salmonella pullorum, and a kit. The primer comprises SP-F and SP-R for detecting the target sequence of the Salmonella pullorum (SP) citE2 gene, and the probe is modified with a fluorescent reporter group and a fluorescent quenching group. By means of providing the primer pair of SP-F and SP-R, which pair specifically binds to the target sequence of the citE2 gene, and combining the primer pair with the probe, whic is used for specific recognition, the accurate and sensitive detection of Salmonella pullorum can be realized.

Graphene electrode for detecting food-borne pathogenic bacteria, sensor and application

Resumen de: CN121142033A

The invention provides a graphene electrode for detecting food-borne pathogenic bacteria, a sensor and application, and belongs to the technical field of analysis of electrochemical sensing. A defect-rich graphene conductive layer is created on the surface of a porous polymer membrane through laser irradiation, then carboxyl-rich magnetic nanoparticles are modified into recognition elements (immunomagnetic beads and IMBs) for capturing and enriching target bacteria in a sample, uncombined free IMBs are separated through nano-filtration of a membrane chip, and the target bacteria in the sample are detected. The larger IMBs-bacterial compound is retained, and the IMBs-bacterial compound retained on the LIG layer confinement structure generates impedance signal change in the micro-electrochemical cell, so that bacterial detection is realized. Meanwhile, the PES-LIG electrode provided by the invention can highly sensitively identify trace target bacteria in a complex matrix.

Primer probe combination, kit and detection method for detecting respiratory tract pathogenic bacteria

Resumen de: CN121137198A

The invention provides a primer probe combination for detecting respiratory tract pathogenic bacteria. The primer probe combination comprises a primer probe combination for detecting klebsiella pneumoniae, a primer probe combination for detecting staphylococcus aureus, a primer probe combination for detecting pseudomonas aeruginosa and a primer probe combination for detecting escherichia coli. The invention also provides a method for detecting respiratory tract pathogenic bacteria by using the primer probe combination. The method comprises the following steps: extracting DNA of a respiratory tract sample; respectively carrying out micro-droplet generation, PCR (Polymerase Chain Reaction) amplification and micro-droplet detection on the extracted respiratory tract sample DNA by using the primer probe combination for detecting the respiratory tract pathogenic bacteria; and interpreting the result. By optimizing the primer design and the PCR amplification system, the kit has the advantages of high specificity, high sensitivity, wide detection range and the like, is rapid and accurate in detection, does not need microbiological culture bacterium expansion on a sample, and is simple and convenient to operate, high in detection flux and short in whole-process detection time.

Kit for rapidly detecting five calf diarrhea related pathogenic microorganisms

Resumen de: CN121142040A

The invention provides a kit for rapidly detecting five calf diarrhea related pathogenic microorganisms, which comprises a colloidal gold test strip for ELISA (enzyme-linked immuno sorbent assay) antigen detection, and a monoclonal antibody is dotted on the test strip. Wherein the monoclonal antibody is prepared by immunizing mice with antigen proteins of bovine rotavirus, bovine coronavirus, escherichia coli, cryptosporidium and clostridium perfringens. The kit for rapidly detecting five pathogenic microorganisms related to calf diarrhea provided by the invention has high detection accuracy, can simultaneously detect five pathogenic microorganisms causing calf diarrhea, and does not cause pathogenic microorganism pollution and affect biological safety. And field operation can be performed, detection is performed immediately after sampling, and a result is given in a relatively short time. The kit disclosed by the invention is simple in detection operation and low in requirements on personnel and equipment.

Rapid immunochromatography detection kit for gastrointestinal pathogenic bacteria based on viral nano-enzyme probe

Resumen de: CN121142029A

The invention belongs to the technical field of biological detection, and discloses a rapid immunochromatography detection kit for gastrointestinal pathogenic bacteria based on a viral nano-enzyme probe. The invention discloses a nano-enzyme probe, the virus-shaped nano-enzyme probe provided by the invention takes Fe3O4 nano-particles as a core, Pt nano-particles and Au (at) Pt nano-particles are coated in sequence, a specific antibody is modified, and the nano-enzyme probe has a magnetic enrichment effect and a multi-catalytic-site characteristic at the same time; rapid capture and high-sensitivity quantitative detection of various gastrointestinal pathogenic bacteria in complex matrixes (river water samples, lake water samples and excrement samples) can be realized, and matrix interference in practical application is eliminated; and the kit has huge potential in the aspects of high-sensitivity field screening and clinical monitoring of infection of gastrointestinal bacteria (such as escherichia coli, salmonella and helicobacter pylori).

LAMP (loop-mediated isothermal amplification) primer group, kit and detection method for detecting pathogenic bacteria of oat smut

Resumen de: CN121137253A

The invention belongs to the technical field of plant fungus molecular biology detection, and discloses an LAMP primer group, a kit and a detection method for detecting oat smut pathogenic bacteria. A group of LAMP (loop-mediated isothermal amplification) specific primers are designed according to a specific sequence on a whole genome of the oat smut pathogenic bacteria Ustilago hordei, results are judged through a real-time fluorescence quantification method, an agarose gel electrophoresis method and an SYBR Green I fluorescent dye developing method, and the oat smut pathogenic bacteria carried by oat seeds are detected. The LAMP detection method for the oat smut pathogenic bacteria, established by the invention, is strong in specificity, high in sensitivity, high in speed and low in cost, provides a new technical means for detection of the oat smut pathogenic bacteria carried by the oat seeds, and has relatively high practical application value.

Pseudomonas aeruginosa InA protein and application thereof

Resumen de: CN121135834A

The invention discloses a pseudomonas aeruginosa InA protein and application thereof, and relates to the technical field of biological detection, the InA protein is InAS2, InAAP41 or InAS8, the amino acid sequences of the InA protein are respectively shown as SEQ ID NO.1-3, and the nucleotide sequences for coding the InAS2, the InAAP41 and the InAS8 are shown as SEQ ID NO.4-6. Compared with the prior art, the InA protein can improve the solubility of target protein, in addition, the InA protein and pseudomonas aeruginosa immune protein also have ultrahigh affinity, a biosensing chip with regeneration reversibility is developed based on the InA protein, the chip can capture fusion protein with the InA protein, and the biosensing chip has the advantages that the biosensing chip can be used as a biosensing chip with regeneration reversibility, so that the biosensing chip can be applied to biosensing of pseudomonas aeruginosa. The chip can be used for detecting the affinity between the fusion protein with the InA tag and a candidate drug, drug screening is realized, and meanwhile, the protein-drug affinity kinetics detected by the chip can also be used for pharmacological research of drugs.

Porcine delta coronavirus N protein monoclonal antibody and fluorescence immunochromatography detection kit and application thereof

Resumen de: CN121135871A

The invention relates to a porcine delta coronavirus N protein monoclonal antibody as well as a fluorescence immunochromatography detection kit and application thereof. According to the application, a prokaryotic expression system is utilized to express the N protein in escherichia coli, the purified N protein is taken as an immunogen, a BALB/c mouse is immunized by an immunological method to prepare the anti-PDCoV N protein monoclonal antibody with high affinity and high potency, and the antibody can be applied to preparation of a PDCoV detection reagent or vaccine. Meanwhile, the invention provides a PDCoV antibody detection kit, N protein and quantum dots (QDs) are subjected to covalent coupling through an EDC method, a fluorescent probe (QDs-Ag) is synthesized, and PDCoV antibody detection is conducted on test paper sprayed with a T line (SPA) and a C line (mAb). The specificity is good, the sensitivity is high, and convenience and accuracy are both considered.

Method for simultaneously extracting and detecting nucleic acid of listeria monocytogenes and salmonella and application

Resumen de: CN121109557A

The invention belongs to the technical field of food safety detection, and relates to a method for simultaneously extracting and detecting nucleic acid of listeria monocytogenes and salmonella. The method comprises the following steps: cracking listeria monocytogenes and salmonella by using a bacterial lysis solution and releasing nucleic acid of listeria monocytogenes and salmonella; the nucleic acid of the listeria monocytogenes and the nucleic acid of the salmonella are extracted at the same time through the synthesized magnetic beads Fe3O4 and Al < 3 + >, and the nucleic acid of the listeria monocytogenes and the nucleic acid of the salmonella are detected at the same time through double RPA/RT-RPA and RPA-LFA test paper. The method has the characteristics of rapidness, high sensitivity and low equipment requirement, and has practical application value in on-site instant detection of food safety.

Rapid detection method for bloodstream infection pathogenic bacteria based on rapid mass spectrometry technology

Resumen de: CN121122425A

The invention discloses a rapid detection method for bloodstream infection pathogenic bacteria based on a rapid mass spectrometry technology, and belongs to the technical field of medical detection. Analyzing the metabolic gas of the blood flow infected pathogenic bacteria by using UVP-TOFMS to obtain a pathogenic bacteria metabolic profile spectrogram; preprocessing the obtained pathogenic bacterium metabolism profile spectrogram data; integrating the preprocessed pathogenic bacterium metabolism profile spectrogram data by using a stacking algorithm, and constructing a pathogenic bacterium identification model; evaluating the effectiveness of the pathogenic bacterium identification model on a training set and a test set through accuracy, precision, a confusion matrix and a subject operation curve; and comparing and screening characteristic metabolic markers of the pathogenic bacteria by combining the importance and significance of model characteristics, and indicating the types of the pathogenic bacteria. The method does not need sample pretreatment, is simple and convenient to operate, short in detection time (20 seconds per sample), high in accuracy and suitable for clinical popularization, and provides technical support for early diagnosis and treatment of bloodstream infection.

Method for rapidly detecting viable salmonella and application

Resumen de: CN121109656A

The invention belongs to the technical field of microbiological detection, and particularly relates to a method for rapidly detecting viable salmonella and application. The method comprises the following steps: mixing a sample to be detected with Felix O-1 bacteriophage, incubating at 37 DEG C to enable the Felix O-1 bacteriophage to infect and proliferate in the sample to be detected, and performing splitting decomposition treatment after infection proliferation is finished to obtain proliferated Felix O-1 bacteriophage DNA (Deoxyribose Nucleic Acid); felix O-1 bacteriophage DNA is used as a template for real-time fluorescence RPA amplification, and whether viable salmonella exists or not is judged through fluorescence signal intensity. According to the method provided by the invention, the defects that the traditional bacterial culture and biochemical confirmation method is long in time consumption and the original molecular biological method cannot conveniently distinguish dead and live are overcome. The method is short in detection time, the RPA reaction is carried out under a constant-temperature condition, equipment requirements are simple, and the method is suitable for field detection.

Listeria monocytogenes detection system based on split G quadruplex cascade CRISPR/Cas12a system and application

Resumen de: CN121109619A

The invention discloses a listeria monocytogenes detection system based on a split G-quadruplex cascade CRISPR/Cas12a system and application of the listeria monocytogenes detection system, and belongs to the technical field of food safety detection. According to the detection method, a split G quadruplex probe is applied, and the split G quadruplex probe is composed of three incomplete guanine-rich DNA chains, namely a G-a chain, a G-b chain and a Linker chain. When no target exists, the CRISPR/Cas12a system is not activated, the Linker chain is complete, and a complete G quadruplex is assembled to be combined with the fluorescent dye to emit fluorescence; when a target exists, an RPA amplification product activation system reversely cuts a Linker chain, a split-chain G quadruplex fails to assemble, ThT cannot be embedded, a fluorescence signal is remarkably reduced, the split G quadruplex is used for replacing a traditional fluorescence modified probe, the method has the advantages of being low in cost, high in sensitivity, high in specificity and the like, and a novel technical platform is provided for detecting listeria monocytogenes in food.

Receptor binding protein for beta-lactam antibiotic detection

Resumen de: CN121086037A

The invention provides a receptor binding protein for beta-lactam antibiotic detection. According to the present invention, efficient soluble expression is performed on the blaRCT gene derived from Bacillus licheniformis in Escherichia coli, such that the receptor binding protein blaRCT capable of being used for beta-lactam antibiotic detection is obtained; detection results show that the receptor binding protein blaRCT has very high affinity and sensitivity to various beta-lactam antibiotics, and the stability of the receptor binding protein blaRCT is still kept above 80% after the receptor binding protein blaRCT is placed at 4 DEG C for half a year. The receptor binding protein has the advantages of high affinity, high sensitivity and high stability, and lays a foundation for subsequent research of beta-lactam antibiotic receptor binding proteins.

Method for detecting five virulence factors of enterotoxigenic escherichia coli in pig farm based on 5-plex mPCR

Resumen de: CN121065316A

The invention provides a method for detecting five virulence factors of enterotoxigenic escherichia coli in a pig farm based on 5-plex mPCR, and belongs to the technical field of biological detection. The multiplex PCR primer group comprises a primer pair used for detecting escherichia coli STa fimbriae genes, a primer pair used for detecting escherichia coli STb fimbriae genes, a primer pair used for detecting escherichia coli LTb fimbriae genes, a primer pair used for detecting escherichia coli STX2e fimbriae genes and a primer pair used for detecting escherichia coli EAST1 fimbriae genes. The invention provides a method for detecting five virulence factors of enterotoxigenic escherichia coli in a pig farm based on 5-plex mPCR. The aim of simultaneously, accurately and efficiently detecting the five most critical virulence factors (STa, STb, LTb subunit, STX2e and EAST1) of swine ETEC is achieved.

Rapid ratio colorimetric detection method for salmonella typhimurium based on hybridization chain reaction

Resumen de: CN121065371A

The invention provides a salmonella rapid ratio colorimetric detection method based on a hybridization chain reaction, and relates to the technical field of biosensing and nucleic acid detection, and the salmonella is adsorbed and enriched by Fe3O4 (at) PDA (at) PEI magnetic nanoparticles through electrostatic interaction. A salmonella specific hairpin aptamer is used for identifying and triggering a hybridization chain reaction (HCR), and a DNA sequence connected with urease is introduced as a signal detection unit. Phenol red is used as an indicator, and the absorbance and the solution color change in a ratio mode. The kit is rapid, sensitive, specific and easy to use, the sample treatment process is reduced, and the sample detection efficiency is improved; the method can be directly operated at room temperature without complex operation environment, and is convenient to use.

Double probes for detecting salmonella, kit and preparation method and application thereof

Resumen de: CN121068916A

The invention provides double probes for detecting salmonella, a kit as well as a preparation method and application of the kit, and belongs to the technical field of bacterial detection. The double probes comprise a capture probe and a detection probe; the capture probe is a magnetic bead coupled with a salmonella aptamer and a salmonella bacteriophage, and the detection probe is gold nanoparticles modified with fDNA; the salmonella aptamer and the fDNA are of a single-chain structure with mutually paired sequences. The double probes provided by the invention respectively promote the sensitive and convenient development of a salmonella detection technology through efficient capture, separation and enrichment and visual signal output. The scheme provided by the invention is simple to operate, low in cost, less in time consumption, easy to implement, high in sensitivity and high in accuracy, does not need large experimental equipment, and can realize visualization of detection results.

Multifunctional Escherichia coli probe protected by silicon shell, preparation method and application of multifunctional Escherichia coli probe in inflammatory marker detection

Resumen de: CN121049496A

The invention discloses a silicon shell protected Escherichia coli multifunctional probe, a preparation method and application of the probe in inflammatory marker detection, the surface of Escherichia coli is sequentially coated with a layer of gold nanoparticles (AuNPs), a layer of silicon dioxide (SiO2) and two layers of quantum dots (QDs), and rapid monitoring of sepsis biomarkers is achieved. The SiO2 layer introduced into the EASi-QDs structure has dual functions of remarkably enhancing the structural stability of bacterial cells and effectively inhibiting the fluorescence quenching effect caused by Au NPs, so that the high sensitivity and accuracy of lateral flow immunochromatography (LFIA) detection are ensured. The EASi-QDs-LFIA has the functions of colorimetric qualitative detection and fluorescent quantitative analysis, and can be used for synchronously detecting two key sepsis markers, namely procalcitonin (PCT) and interleukin-6 (IL-6). The detection limits of the method on PCT and IL-6 respectively reach 29.5 pg/mL and 4.7 pg/mL, which are respectively improved by at least 15 times and 339 times compared with the traditional enzyme-linked immunosorbent assay (ELISA) and gold nano colorimetric method.

Preparation method, product and application of quality control product for pathologic detection of pathogenic bacteria

Resumen de: CN121046554A

The invention discloses a preparation method of a pathologic detection quality control product for pathogenic bacteria, which comprises the following steps: mixing target pathogenic bacteria and eukaryotic cells with curable hydrogel according to a required ratio to form a bacteria-containing biological matrix, and curing and forming by using a three-dimensional forming method to form a three-dimensional solid structure; the pathologic detection quality control product containing the microorganism-cell composite structure of the simulated pathological tissue is obtained. The preparation process is stable and controllable, uniform distribution of pathogenic bacteria is guaranteed, ethical problems do not exist, and clinical product transformation can be rapidly carried out. The invention further discloses a product obtained through the preparation method and application of the product. The sample obtained by the method can be used as a standardized positive control sample in the whole pathologic detection process of pathogenic bacteria, and is used for quality control. The invention solves the problem of lack of full-process quality control products in existing pathogenic bacterium detection, and is suitable for full-process quality control of tissue block preparation, pathological section dyeing, molecular detection and the like.

Multiple qPCR (quantitative polymerase chain reaction) detection kit for simultaneously detecting four fimbriae genes of swine pathogenic escherichia coli and application of multiple qPCR detection kit

Resumen de: CN121023062A

The invention discloses a multiple qPCR (quantitative polymerase chain reaction) detection kit for simultaneously detecting four fimbriae genes of swine pathogenic escherichia coli and application of the multiple qPCR detection kit. The kit comprises primers and a TaqMan probe, wherein the primers and the TaqMan probe are respectively used for detecting F4, F17, F18 and F41 fimbriae genes of swine pathogenic escherichia coli. Experiments prove that the kit has the advantages of low cost, high specificity, high sensitivity, rapidness and reliability in detection, simplicity in operation and the like when being used for detection. According to recent epidemiological investigation results, 951 pig anus swab samples are detected by using the method, 662 positive samples are detected, the positive rate is 69.61%, the sensitivity of the method is higher than that of conventional PCR, and the total coincidence rate of the two results is greater than 90%. The method is suitable for rapid molecular diagnosis of swine pathogenic Escherichia coli, is an effective technical means for screening the virulence gene of the swine pathogenic Escherichia coli epidemic strain, and fills the blank of multiple fluorescent PCR detection technology in the field.

Avian salmonella nucleic acid rapid detection method and kit based on RAA-CRISPR-Cas13a technology

Resumen de: CN121023055A

The invention discloses an avian salmonella nucleic acid rapid detection method based on a RAA-CRISPR-Cas13a technology and a kit, and belongs to the technical field of biochemistry and molecular biology. On the basis of the designed RAA primer pair and crRNA, the avian salmonella nucleic acid detection based on an RAA-CRISPR-Cas13a enzyme molecular system is established, the system is high in specificity and sensitivity, rapid detection of avian salmonella can be achieved within 30 minutes, the detection sensitivity of a fluorescence method can reach 100 copies/microliter, and the detection sensitivity of a test paper method can reach 102 copies/microliter. In addition, according to the system, expensive test equipment does not need to be operated in a standard professional laboratory, and field detection of clinical samples can be achieved through the portable visual transverse flow test strip.

Method for detecting activity and concentration of Candidatus Liberobacter asiaticum pathogenic bacteria in plant tissue

Resumen de: CN121027060A

The invention discloses a method for detecting the activity and concentration of Candidatus Liberobacter asiaticum pathogenic bacteria in plant tissues, relates to the technical field of microbiological detection, and is used for rapidly detecting the activity of the Candidatus Liberobacter asiaticum pathogenic bacteria in Citrus and related plant tissues infected with Citrus Liberobacter asiaticum through flow cytometry. Live/dead cells of the CLas can be visually distinguished, the concentration of the CLas in plant tissues is obtained, and detection can be completed within 30 minutes at the soonest. Compared with conventional fluorescent quantitative PCR detection which needs complex operation of multiple steps, the method has the advantages that the detection time is long, and the operation of the flow cytometer is more convenient and faster. More importantly, the method disclosed by the invention can realize real-time distinguishing of living cells and dead cells of the CLas, while the fluorescent quantitative PCR method is used for detecting DNA in the cells, and the physiological status of the cells cannot be judged. As dead cells still retain undegraded DNA within a certain period of time, the continuous existence of a fluorescence signal causes that the detection result of the concentration of pathogenic bacteria lags behind the actual viable bacteria level.

LOOP-MEDIATED ISOTHERMAL AMPLIFICATION PRIMERS FOR SHIGA TOXIN-PRODUCING E. COLI (STEC) DETECTION

Nº publicación: US2025361570A1 27/11/2025

Solicitante:

NEOGEN FOOD SAFETY US HOLDCO CORP [US]
NEOGEN FOOD SAFETY US HOLDCO CORPORATION

Resumen de: US2025361570A1

Primers for of amplifying DNA from shiga toxin producing E. coli (STEC); methods of amplifying STEC DNA; and methods of detecting STEC.