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Reverse transcription real-time quantitative polymerase chain reaction (RT-qPCR) detection method aiming at escherichia coli total ribonucleic acid (RNA) residues and application

Resumen de: CN122081523A

The invention belongs to the technical field of biological detection, and particularly relates to a high-sensitivity and high-specificity quantitative detection method for total RNA (ribonucleic acid) of Escherichia coli remained in the production process of biological products (such as recombinant proteins, vaccines, cell therapy products and the like). Particularly relates to detection system construction, primer probe design and standardized operation processes based on a reverse transcription real-time quantitative polymerase chain reaction (RT-qPCR) technology, and further expands application of the detection system construction, primer probe design and standardized operation processes in drug quality control, process verification and cleaning verification.

Rapid and accurate detection method for pathogenic bacteria

Resumen de: CN122060834A

The invention discloses a rapid and accurate detection method for pathogenic bacteria. The method comprises the following steps: immobilizing bacteriophages on the surfaces of magnetic nanoparticles through physical adsorption or chemical reaction, selectively enriching pathogenic bacteria in a complex sample by adopting the bacteriophage functionalized magnetic nanoparticles, quickly cracking the pathogenic bacteria on the surfaces of the magnetic nanoparticles by using CTAB (cetyltrimethyl ammonium bromide), releasing ATP in the bacteria, and quantitatively analyzing ATP by using bioluminescence. Further calculating the content of the pathogenic bacteria; the detection method is free of enrichment and amplification, can complete capture, enrichment and detection of pathogenic bacteria within 20 minutes, and can specifically detect viable bacteria of target bacteria in various complex matrix (such as food) samples due to the fact that bacteriophages are adopted as recognition molecules.

Kit and detection method for detecting multiple piglet diarrhea pathogens

Resumen de: CN122060931A

The invention provides a kit for detecting multiple piglet diarrhea pathogens and a detection method, and belongs to the technical field of biological information detection. The kit comprises a reaction reagent, wherein the reaction reagent comprises a porcine epidemic diarrhea virus specific primer and a probe, a porcine delta coronavirus specific primer and a probe, a piglet pathogenic escherichia coli specific primer and a probe, a clostridium perfringens specific primer and a probe, a buffer solution, an enzyme mixed solution and a freeze-drying protective additive. The detection method adopts the kit for detecting various piglet diarrhea pathogens. According to the kit disclosed by the invention, simultaneous detection of four target genes including the porcine epidemic diarrhea virus, the porcine delta coronavirus, the piglet pathogenic escherichia coli and the clostridium perfringens can be realized only through one-time amplification, so that the detection time is greatly shortened while the detection cost is reduced; the kit has extremely high specificity, sensitivity and repeatability, and the primers and the probes do not interfere with each other; good normal-temperature storage stability is also realized.

Holding type listeria monocytogenes specific molecular target and rapid detection method thereof

Resumen de: CN122060880A

The invention discloses a persistent listeria monocytogenes specific molecular target and a rapid detection method thereof, and belongs to the technical field of microbiological detection. The molecular target is obtained through bioinformatics analysis and screening, is further screened through specific experimental verification, and is respectively specific genome sequence fragments of six retention types of listeria monocytogenes CC5, CC8, CC31, CC87, CC121 and CC204. A corresponding specific primer group is designed based on the target, sample DNA is amplified through PCR, a product is subjected to visual analysis in combination with gel electrophoresis, and whether the target retention type strain exists or not can be judged according to the size of a band. The detection method has the advantages of being short in detection time, low in cost, simple and convenient to operate and high in specificity, results are accurate, judgment is easy, practicability is high, the limitation that traditional culture and identification periods are long, and phenotypic analysis steps are tedious is broken through, and the detection method is suitable for rapid screening of the persistent listeria monocytogenes in samples of food, environment and the like.

Au-TiO2 coating-based pathogenic bacteria splitting decomposition and hydrogel in-situ detection method

Resumen de: CN122060843A

The invention discloses a pathogenic bacteria splitting decomposition and hydrogel in-situ detection method based on an Au-TiO2 coating, and belongs to the technical field of biology. The method comprises the following steps: (1) preparing an Au-TiO2 composite coating on a substrate; (2) dropwise adding a to-be-detected liquid sample to the surface of the Au-TiO2 composite coating, and covering the hydrogel sheet after the solvent is evaporated; or adhering the hydrogel sheet to the surface of the solid sample to be detected, stripping the adsorbed pathogenic bacteria, and covering the Au-TiO2 composite coating with the hydrogel sheet; the hydrogel sheet contains lysozyme, an isothermal amplification reaction reagent and a fluorescent dye; and (3) incubating the reaction system under an isothermal amplification reaction condition, and carrying out fluorescence imaging after the reaction is finished. According to the method provided by the invention, through hydrogel fixation and synergistic effect of lysozyme and Au-TiO2, splitting decomposition of pathogenic bacteria on an interface layer is promoted, in-situ nucleic acid amplification is realized, and the method has an extremely high application value for on-site rapid detection of pathogenic bacteria of agricultural products.

Primer group, kit and detection method for detecting salmonella enteritidis

Nº publicación: CN122060881A 19/05/2026

Solicitante:

NINGXIA HUI AUTONOMOUS REGION FOOD INSPECTION AND RESEARCH INST
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Resumen de: CN122060881A

The invention belongs to the technical field of microbiological detection and molecular diagnosis, and discloses a primer group, a kit and a detection method for detecting salmonella enteritidis, the kit comprises an immunocapture component, an isothermal amplification component, a CRISPR/Cas12a detection component and auxiliary components; the isothermal amplification component is any one of an RPA (recombinase polymerase amplification) component or an LAMP (loop-mediated isothermal amplification) component, the RPA component comprises a specific primer group targeting 86th to 212th nucleotides in a salmonella enteritidis hilA gene coding region, and the LAMP component comprises a specific primer group targeting 58th to 250th nucleotides in the salmonella enteritidis hilA gene coding region. The primer group, the kit and the detection method for detecting the salmonella enteritidis successfully integrate three key technologies of RAP and LAMP rapid isothermal amplification, CRISPR/Cas12a high-specificity recognition and trans-cleavage and immunomagnetic bead targeted enrichment, and creatively provide the primer group, the kit and the detection method for detecting the salmonella enteritidis.