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Synchronous detection method for klebsiella pneumoniae and escherichia coli based on RPA-CRISPR/Cas12a and application

Resumen de: CN121272079A

The invention relates to the technical field of molecular biology and pathogenic microorganism detection, and discloses a Klebsiella pneumoniae and Escherichia coli synchronous detection method based on RPA-CRISPR/Cas12a and application, and the method comprises the following steps: extracting DNA from a sample, carrying out isothermal amplification by adopting an RPA primer designed aiming at a Klebsiella pneumoniae rcsA gene and an Escherichia coli uriA gene, and carrying out amplification by adopting an RPA primer designed aiming at a Klebsiella pneumoniae RcsA gene and an Escherichia coli uriA gene; a detection system containing Cas12a enzyme, crRNA and a fluorescence report substrate is constructed, and a result is interpreted through a fluorescence or lateral flow chromatography test strip. Detection can be completed within 70 minutes at the constant temperature of 37 DEG C, the sensitivity reaches 5 * 10 copies/mu L, the specificity is high, no cross reaction exists, the kit is suitable for clinical early screening, hospital infection monitoring and on-site POCT application, and an efficient and reliable new tool is provided for pathogen diagnosis.

C8 esterase activated double-lock chemiluminescent probe, preparation method thereof and application of C8 esterase activated double-lock chemiluminescent probe in preparation of detection kit for detecting salmonella

Resumen de: CN121270430A

The invention relates to the technical field of chemical analysis and detection, and discloses a C8 esterase activated double-lock chemiluminescence probe, a preparation method thereof and application of the C8 esterase activated double-lock chemiluminescence probe in preparation of a salmonella detection kit, the double-lock chemiluminescence probe is DSCP-CBCN, and the structural formula of the double-lock chemiluminescence probe is shown in the specification. The DSCP-CBCN probe is composed of a C8 esterase cleavable substrate, a phenoxy cyclobutane chemiluminophor and a self-eliminating connexon p-hydroxy benzyl alcohol. After the DSCP-CBCN probe and salmonella are incubated, sodium hypochlorite and hydrogen peroxide are added to generate remarkable chemiluminescence, POCT rapid detection of salmonella can be realized by combining a chemiluminescence microplate reader, the bacterial culture time is shortened to 3-7 hours, the limit of detection (LOD) value is as low as 1.9 CFU/mL, and the sensitivity is improved by 1008 times compared with that of a commercial MUCAP probe. The method is suitable for salmonella large-scale screening in the fields of clinical pathogen diagnosis, food safety detection and environmental health monitoring.

Electrochemical sensing detection system and method based on bacterial nucleotide metabolism

Resumen de: CN121253627A

The invention relates to the technical field of water toxicity detection, in particular to an electrochemical sensing detection system and method based on bacterial nucleotide metabolism. Comprising the following steps: dispersing carboxylated multi-walled carbon nanotubes in N, N-dimethylformamide to prepare a dispersion liquid with a concentration of 0.5 to 1 mg/mL; adding ZnO nanoparticles, and performing ultrasonic treatment to obtain a mixed dispersion liquid; and dispensing the mixture on the surface of a pretreated glassy carbon electrode, and drying the glassy carbon electrode under an infrared lamp to obtain the nano-composite modified electrode. A three-electrode system formed by taking the nano-composite modified electrode as a working electrode, taking saturated calomel as a reference electrode and taking a platinum column as a counter electrode is connected to an electrochemical workstation; a guanine and xanthine mixed oxidation peak current generated by metabolism of Escherichia coli is used as a comprehensive toxicity effect index, and the toxicity of a target system is evaluated by monitoring the change of the mixed oxidation peak current. According to the invention, by utilizing the synergistic interaction of the nano composite material, the sensitive and rapid detection of the comprehensive toxicity of the water body is realized without adding a mediator.

Method and device for quantitatively detecting pathogenic bacteria and detecting sensitivity of antibacterial agent

Resumen de: CN121249844A

The invention relates to the technical field of biosensing, in particular to a pathogenic bacteria quantitative detection and antibacterial drug sensitivity detection method and device. Based on the relationship between the concentration of gelatin and the time when a to-be-detected solution passes through a detection interval from bottom to top, a pathogenic bacteria quantitative detection and antibacterial drug sensitivity detection method is obtained; according to the quantitative detection method for the pathogenic bacteria, on the basis of ensuring the sensitivity, the detection time is shortened, and staphylococcus aureus as low as 1 CFU/mL can be detected after incubation for 8 hours. The MIC value measured by the antibacterial drug sensitivity detection method is highly consistent with the result of the traditional gold standard method (broth microdilution method), but the required time is greatly shortened (only about 6 hours are needed, and the standard method needs 18-24 hours).

PCR kit for triple PCR detection of escherichia coli, salmonella enteritidis and riemerella anatipestifer and application of PCR kit

Resumen de: CN121249921A

The invention belongs to the technical field of triple PCR detection PCR kits, and particularly relates to a triple PCR detection PCR kit for escherichia coli, salmonella enteritidis and riemerella anatipestifer and application of the triple PCR detection PCR kit. According to the kit, specific primers are designed by taking an escherichia coli alkaline phosphatase gene, a salmonella enteritidis invasion protein A gene and a riemerella anatipestifer helicase gene as target genes, the specific primers comprise a primer pair aiming at the phoA gene, the sequence of an upstream primer phoA-F is 5 '-TACAGGTACTGCGGGCTTATC-3', the sequence of a downstream primer phoA-R is 5 '-CTTACCGGGAATACATCACA-3', and the length of an amplified target fragment is 622bp; according to the primer pair for the invA gene, the sequence of an upstream primer invA-F is 5 '-AAAAGAAGGGTCGTCGTTAG-3', the sequence of a downstream primer invA-R is 5 '-GGAAGGTACTGCCAGAGGTC-3', the length of an amplified target fragment is 801 bp, and according to the primer pair for the dnaB gene, the sequence of an upstream primer dnaB-F is 5 '-GACGCTATATCATAGATTTAAC-3', the sequence of a downstream primer dnaB-R is 5 '-TCCACCGCTATATATTCTAG-3', and the length of the amplified target fragment is 431 bp.

Detection system for rapidly identifying drug resistance phenotypes of pathogenic bacteria based on machine learning in combination with MALDI-TOF MS (matrix-assisted laser desorption ionization-time of flight mass spectrometry)

Resumen de: CN121237227A

The invention relates to the technical field of clinical microbiological detection, in particular to a detection system for rapidly identifying drug resistance phenotypes of pathogenic bacteria based on machine learning in combination with MALDI-TOF MS. According to the system, optimal detection conditions are screened out, clinically separated pathogenic bacteria and antibiotics are subjected to grouping co-incubation, and MALDI-TOF MS detection is carried out through dynamic sampling at multiple time points; preprocessing the obtained mass spectrum data, and calculating to obtain an enhanced dynamic relative growth (RBD-RG) feature vector matrix; a plurality of machine learning models are trained based on the matrix, and an optimal drug resistance phenotype prediction model is screened out through cross validation, so that the drug resistance phenotypes (sensitivity, mediation and drug resistance) of pathogenic bacteria are quickly and accurately judged. The whole detection process can be completed within 2 hours, the detection efficiency is remarkably improved, key technical support is provided for early-stage precise medication of infectious diseases, and improvement of prognosis of patients and restraint of infection spreading are facilitated.

Method for detecting mouse salmonella by using qPCR (quantitative polymerase chain reaction) technology instead of traditional sacrificial sentinel mouse

Resumen de: CN121227909A

The invention discloses a method for detecting mouse salmonella by using a qPCR (quantitative polymerase chain reaction) technology instead of a traditional sacrificial sentinel mouse, and a salmonella specific primer and probe combination comprises a forward primer with a sequence of 5 '-AGCGTACTGGAAAGGGAAAG-3', as shown in SEQ ID NO.1, a reverse primer with a sequence of 5 '-AGCGTACGAAAG-3', as shown in SEQ ID NO.2, a reverse primer with a sequence of 5 '-AGCGTACGAAGGAAAG-3', as shown in SEQ ID NO.3, the sequence of a reverse primer is 5-ATACCGCCAATAAAGTTCACAAAG-3 ', and the sequence of the reverse primer is as shown in SEQ ID NO. 2; the probe sequence of the probe is 5 '-FAM-CGTCACCTTGATAAACTCTATGCA-BHQ1-3', and the probe sequence of the probe is as shown in SEQ ID NO. 3. The method has the beneficial effects that a real-time fluorescent quantitative PCR method is established by utilizing the specific primer and the double-labeled fluorescent probe, so that rapid detection of salmonella pollution in facilities is realized; according to the method based on the environmental sample, use of additional animals is avoided, operation is easy and convenient, the result is accurate, and sensitivity and specificity are both superior to those of a traditional sentinel mouse method.

Multiplex PCR (Polymerase Chain Reaction) method for rapidly detecting pathogenic bacteria of tobacco brown spot

Resumen de: CN121227938A

The invention discloses a multiplex PCR (polymerase chain reaction) method for quickly detecting pathogenic bacteria of tobacco brown spot, which is used for synchronously identifying two main pathogenic bacteria causing tobacco brown spot, namely alternaria tenuissima and alternaria alternata, and comprises the following steps: S1, extracting genome DNA (deoxyribonucleic acid): extracting genome DNA of standard strains of alternaria tenuissima and alternaria alternata as a detection template; the multiple PCR system has the beneficial effects that through system verification, it is confirmed that all primers only generate expected amplification products for target pathogenic bacteria, no cross amplification reaction exists between alternaria alternata and alternaria tenuissima, and it is indicated that the multiple PCR system has high interspecific specificity. The established detection method is high in stability, good in repeatability and suitable for standardized operation under different laboratory conditions. Compared with a traditional morphological identification or single gene sequencing method, the multiplex PCR technology has the advantages that the detection period is remarkably shortened, the operation is simple and convenient, time and labor are saved, the cost is low and the like. The method is suitable for rapid identification of standard strains.

Method for synchronously detecting food-borne pathogenic bacteria based on multiple bacteriophage modified nano-enzyme

Resumen de: CN121231460A

The invention discloses a method for synchronously detecting food-borne pathogenic bacteria based on multiple bacteriophage modified nano enzyme, and belongs to the technical field of biology, a phage coated FeCoCe NF + TMB color development system is adopted to react with a solution to be detected, and the food-borne pathogenic bacteria in the solution to be detected are detected through the color development reaction; in the phage (at) FeCoCe NF + TMB color development system, a preparation method of the phage (at) FeCoCe NF comprises the following steps: immobilizing phages EhpYZU20, SalmpYZU54 and SapYZUH5 on a FeCoCe NF nano enzyme, so as to obtain the phage (at) FeCoCe NF. The preparation method of the phage (at) FeCoCe NF comprises the following steps of: immobilizing phages EhpYZU20, SalmpYZU54 and SapYZUH5 on the FeCoCe NF nano enzyme; according to the invention, synchronous visual detection of three food-borne pathogenic bacteria can be realized.

Detection method and application of mouse pathogenic bacteria

Resumen de: CN121227913A

The invention relates to a multiplex fluorescent quantitative PCR (Polymerase Chain Reaction) method for detecting mouse pathogenic bacteria, which can be used for simultaneously detecting four pathogens, namely salmonella, corynebacterium murine, pseudomonas aeruginosa and klebsiella pneumoniae.

Intestinal escherichia coli gene detection kit based on silicon-based micro-fluidic chip and detection method of intestinal escherichia coli gene detection kit

Resumen de: CN121227912A

The invention discloses an intestinal escherichia coli gene detection kit based on a silicon-based micro-fluidic chip and a detection method of the intestinal escherichia coli gene detection kit. The kit provided by the invention comprises a rapid hot start DNA polymerase, a PCR reaction mixed solution, a positive reference substance and a negative reference substance, the kit also comprises the following primers which take the intestinal escherichia coli fliC gene as a target gene, are designed by a PCR fluorescent probe method based on a micro-fluidic chip technical platform: an upstream primer F, a downstream primer R and a detection probe P, and the nucleotide sequences of the primers are shown as SEQ ID NO.1-3. According to the invention, the intestinal escherichia coli gene detection kit which is more comprehensive in detection effect, high in specificity, good in sensitivity, low in omission ratio, convenient, simple and easy to operate is provided.

Specific primer set of Pseudomonas aeruginosa and method for detecting Pseudomonas aeruginosa using the same

Resumen de: KR20250178279A

본 발명은 서열번호 1의 올리고뉴클레오티드 프라이머 및 서열번호 2의 올리고뉴클레오티드 프라이머를 포함하는, 슈도모나스 에르지노사(Pseudomonas aeruginosa) 검출용 프라이머 세트; 이를 포함하는 조성물; 이를 포함하는 키트; 및 이를 이용한 슈도모나스 에르지노사(Pseudomonas aeruginosa) 검출 방법에 관한 것이다.

Specific primer set of Listeria monocytogenes and method for detecting Listeria monocytogenes using the same

Resumen de: KR20250177874A

본 발명은 서열번호 1의 올리고뉴클레오티드 프라이머 및 서열번호 2의 올리고뉴클레오티드 프라이머를 포함하는, 리스테리아 모노사이토제네스(Listeria monocytogenes) 검출용 프라이머 세트; 이를 포함하는 조성물; 이를 포함하는 키트; 및 이를 이용한 리스테리아 모노사이토제네스(Listeria monocytogenes) 검출 방법에 관한 것이다.

Method for synchronously and rapidly detecting escherichia coli and multi-drug-resistant genes thereof

Resumen de: CN121204211A

The invention provides a method for synchronously and rapidly detecting Escherichia coli and multiple drug-resistant genes thereof, which comprises the following steps: culturing a sample at night to obtain a bacterial solution to be detected, adding a buffer solution and a metal bath, centrifuging, taking a supernatant, detecting by micro-fluidic, optimizing the extraction process of nucleic acid in the sample, designing an optimal RPA primer probe group, and detecting the multiple drug-resistant genes of the Escherichia coli. A reasonable RPA coupled microfluidic detection system is established, synchronous and rapid detection of the escherichia coli uidA and I-type integrase gene intI1 thereof and drug-resistant genes sul1 and aadA5 is realized, the specificity is high, the sensitivity is high, the reaction time is short, the operation is convenient, and a method support is provided for monitoring and prevention and control of food-borne drug-resistant bacteria.

Biosensor for detecting pseudomonas fluorescens and detection method

Resumen de: CN121208335A

The invention belongs to the technical field of food safety and biosensors, and particularly relates to a biosensor for detecting pseudomonas fluorescens and a detection method. The biosensor for detecting the pseudomonas fluorescens, provided by the invention, comprises a nano click enzyme at bacteriophage recognition probe, a magnetic adsorption platform and a click substrate, and is a fluorescent biosensing system based on a click nano enzyme mediated signal amplification strategy and bacteriophage specific recognition. By combining the high catalytic activity of the recognition probe on the click reaction, the high-specificity signal amplification advantage of the recognition probe on a target detection object and the enrichment amplification advantage of magnetic adsorption, the probe shows excellent detection sensitivity and specificity in pseudomonas fluorescens detection, the detection limit is 1 CFU/mL, the linear range is 102-107 CFU/mL, and the detection sensitivity is high. Therefore, technical support can be provided for ultra-sensitive and high-specificity detection of the pseudomonas fluorescens, and the method is very suitable for on-site rapid screening of raw milk, pasteurized milk, UHT milk and diluted samples of the pasteurized milk, the UHT milk and the diluted samples of the pasteurized milk and the UHT milk.

Method for detecting listeria monocytogenes in livestock and poultry meat based on controllable change of peroxidase-like activity

Resumen de: CN121208336A

The invention belongs to the technical field of biological detection, and particularly relates to a method for detecting listeria monocytogenes in livestock and poultry meat based on controllable change of peroxidase-like activity. According to the method, heme, manganese chloride tetrahydrate and sodium hydroxide are taken as substrates, and the nano-enzyme Mn-Hemin with peroxidase-like activity is synthesized through a room-temperature stirring method. The nano-enzyme Mn-Hemin can specifically adsorb and recognize the listeria monocytogenes and regulate and control the peroxidase activity of the listeria monocytogenes, and finally a method for rapidly determining the listeria monocytogenes in a colorimetric mode is established. According to the listeria monocytogenes detection method provided by the invention, the detection time is only 5 minutes. In addition, Mn-Hemin is outstanding in anti-interference performance, various food-borne pathogenic bacteria similar to listeria monocytogenes can be effectively eliminated, the detection limit is 6.56 CFU/mL, the linear range is 1.0-1.0 * 10 < 5 > CFU/mL, and good sensitivity and accuracy are shown.

Primer group for simultaneously detecting four pathogenic microorganisms and application of primer group in preparation of micro-fluidic chip and kit

Resumen de: CN121182993A

The invention discloses a primer group for simultaneously detecting four pathogenic microorganisms and application of the primer group in preparation of a micro-fluidic chip and a kit. The sequences of the primers are as shown in SEQ ID NO: 1-24. The method has the advantages that the microfluidic technology and the isothermal amplification technology are combined, four sample indexes are detected at the same time in one-time detection, and the method has the advantages of being high in throughput, high in specificity, good in experiment repeatability and convenient and rapid to operate; the kit can be used for rapidly and accurately detecting salmonella (SAL), staphylococcus aureus (SA), listeria monocytogenes (LM) and escherichia coli O157: H7 (E. coli O157: H7).

Composition for detecting respiratory tract infection pathogens, kit and application

Resumen de: CN121182992A

The invention relates to the technical field of biological detection, and discloses a composition for detecting respiratory tract infection pathogens, a kit and application. The composition disclosed by the invention can be used for rapidly and simultaneously detecting legionella pneumophila, bordetella pertussis, chlamydia pneumoniae, mycoplasma pneumoniae, mycobacterium tuberculosis, aspergillus and cryptococcus neoformans in a sample with high specificity, and has relatively high sensitivity and stability. Compared with traditional detection methods such as pathogen isolated culture and genome sequencing, the efficiency of identifying and jointly detecting multiple respiratory tract pathogens by adopting the composition disclosed by the invention is effectively improved, the detection process is simple and convenient to operate, and the composition is suitable for large-scale popularization and application.

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;

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.

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.

PSEUDOMONAS AERUGINOSA INA PROTEIN AND USE THEREOF

Nº publicación: WO2025256021A1 18/12/2025

Solicitante:

BIORTUS BIOSCIENCES CO LTD [CN]
BIORTUS DISCOVERY CO LTD [CN]
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\u4F70\u7FF1\u5F97\uFF08\u65E0\u9521\uFF09\u65B0\u836F\u5F00\u53D1\u6709\u9650\u516C\u53F8

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.