Mastitis Pathogen Identification Methods in Raw Milk

Multiplex immunoassay for detecting Mycoplasma bovis infection in cattle using a substrate with multiple capture elements. Each capture element is specific to Mycoplasma bovis antigens, including proteins, lipoproteins, glycoproteins, and peptides, and can bind to target antibodies. The substrate also includes fiduciary markers, control elements, and a colorimetric detection system to monitor assay performance. The assay provides a comprehensive view of Mycoplasma bovis antigens and antibodies in milk or serum samples, enabling accurate detection of the pathogen in clinical samples.

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A method for detecting and monitoring pathogens in food samples using aptamer-based capture or antibody-based capture followed by releasing live pathogens for detection. The method employs a disposable cartridge with a pre-filter container containing bead arrays that capture pathogens from the sample matrix. The captured pathogens are then washed, released, and subsequently detected through nucleic acid amplification, enabling rapid, culture-independent analysis of environmental, food, and water samples.

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A lateral flow assay (LFA) test strip platform for rapid, high-sensitivity detection of pathogens, including viruses and their antigens, using hydrogel-based immobilization of affinity probes. The platform employs a novel hydrogel polymer patterned test strip architecture, where a hydrogel matrix is integrated into the test line structure, enabling controlled immobilization of affinity probes. This architecture allows for precise control over probe orientation and positioning, significantly improving detection sensitivity and specificity compared to conventional LFA methods. The platform can be adapted for detection of a wide range of pathogens, including SARS-CoV-2, and is particularly suitable for rapid, point-of-care diagnostics.

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Disposable cartridge for rapid, low-cost, and portable detection of pathogens like viruses from samples like breath or saliva. The cartridge has separate reservoirs for protein and genetic component buffers, and sensors for detecting the target protein and genetic component. This allows confirming pathogen presence by simultaneous detection of both components. The cartridge can be used with a breath collection device that mixes breath samples with the buffers and introduces them into the sensors.

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A rapid, in-stall method for detecting gram positive mastitis pathogens in bovine milk using luminescence. The method involves: 1) filtering raw milk to remove large particles, 2) rupturing gram positive bacteria in the milk using enzymes like lysostaphin and phage endolysins, 3) eliminating endogenous milk ATP to prevent interference, 4) adding luciferin-luciferase reagent to measure bacterial ATP, and 5) detecting luminescence to quantify gram positive bacteria. The method provides a "cow side" rapid diagnostic for mastitis within 10 minutes using simple reagents and devices that could significantly reduce antibiotic use and costs by enabling targeted treatment of gram positive mastitis.

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A rapid diagnostic kit for mastitis that enables quick identification of pathogenic bacteria causing the disease, while preventing unnecessary antibiotic use. The diagnostic system combines a somatic cell count (SCC) measurement with a unique pathogen-specific fluorescence pattern analysis to provide rapid and accurate identification of mastitis-causing pathogens. The system uses a proprietary fluorescence-based detection method that is more sensitive than traditional somatic cell count methods, allowing for faster and more reliable identification of pathogenic bacteria compared to traditional methods.

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Real-time somatic cell count monitoring system for dairy farms during milking, enabling early detection of mastitis through continuous cell analysis. The system integrates into the milking process, employing a portable, automated somatic cell counter that continuously measures cell counts at various levels along the milking line. This real-time monitoring enables early intervention before clinical mastitis develops, significantly reducing treatment costs and improving herd health. The system operates in parallel with the milking process, with the cell counter positioned between the milk source and the collective tank, allowing continuous testing throughout the milking cycle.

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Devices and assays for detecting foodborne pathogens through rapid, sensitive detection of microbial antigens. The devices comprise a sample inlet, analyte detection cartridge receptacle, and analyte detection cartridge receptacle inlet, along with optional conjugate pad removal and pressure actuation. The device enables the rapid detection of microbial antigens in food samples through a novel membrane-based detection system.

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A nucleotide primer set for simultaneous detection and identification of eight mastitis pathogens through loop-mediated isothermal DNA amplification (LAMP). The primer set enables simultaneous detection of E. coli, Staphylococcus aureus, Klebsiella pneumonia, Streptococcus uberis, Streptococcus dysgalactiae, Streptococcus agalactiae, Mycoplasma bovis, and coagulase-negative Staphylococci (CNS) tuf including subtypes S. epidermidis, S. chromogenes, and S. simulans.

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