Polymerase Chain Reaction Methods for Pathogen Detection in Milk

A real-time quantitative PCR method for detecting Prototheca bovis in dairy products, particularly in non-diagnostic samples. The method employs specific primers targeting the cob gene of Prototheca bovis, which are amplified in a single reaction cycle. The primers are designed to specifically detect the cob gene sequence, eliminating the need for multiple primer pairs and reducing the risk of cross-reactivity. The method provides high sensitivity and specificity for detecting Prototheca bovis in milk and dairy products, enabling accurate quantification of the pathogen in non-diagnostic samples.

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Testing milk for pathogens using a multi-well PCR system that can be performed on non-aseptic milk samples collected through standard dairy herd collection processes. The system enables rapid detection of pathogens in milk using specific PCR reagents and polymerase chain reaction amplification approaches, while maintaining the integrity of the milk samples. This approach can be used for testing milk collected through conventional collection methods, such as DHI, without the need for specialized aseptic collection procedures.

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PCR primer pair for detecting Bacillus sporothorax (BSTD) in dairy products, optimized for quantitative analysis with high specificity. The primer pair is designed to amplify BSTD DNA with minimal cross-reactivity, enabling accurate quantification of BSTD in dairy products. The primer pair is particularly effective in dairy products where BSTD is present at concentrations below 2 million CFU/mL. The detection method utilizes a fluorescence-based PCR approach, allowing for rapid quantification and detection of BSTD in dairy products.

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A real-time quantitative PCR method for detecting multiple bovine mastitis pathogens using a combination of primers and probes. The method employs a unique combination of TaqMan probes that specifically amplify the target genes of various pathogenic bacteria, including Staphylococcus aureus, Streptococcus, Mycoplasma, Staphylococcus chromogens, and Escherichia coli. The probes are designed to target specific sequences in each pathogen, enabling rapid and accurate detection of multiple pathogens simultaneously. The method achieves high sensitivity and specificity, with demonstrated performance in detecting pathogens like S. aureus, S. uberis, M. bovis, Staphylococcus chromogens, and E. coli. The combination of TaqMan real-time PCR with this specific primer/probe combination enables rapid and efficient diagnosis of bovine mastitis, as well as non-diagnostic purposes like laboratory screening and food safety inspection.

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A method for detecting bovine and horse-derived components in raw milk and fermented milk through real-time PCR-based detection using primers, probes, and kits. The method employs a combination of bovine-specific primers and probes, along with horse-derived probes, to simultaneously detect the presence of bovine and horse-derived DNA in milk and fermented products. This approach enables the detection of both bovine and horse-derived components in raw milk and fermented milk samples, particularly in low-probability detection scenarios.

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A rapid detection method for Staphylococcus aureus in milk and raw milk using a PCR-based assay. The method employs a specific primer set targeting the manganese transport protein C gene (MntC) of Staphylococcus aureus, which is highly conserved across strains and serotypes. The primers are designed to amplify the MntC gene in the presence of manganese ions, making it a reliable marker for detecting this critical virulence factor. The assay combines the extraction of microorganisms from milk with the rapid PCR detection of MntC gene sequences.

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A rapid detection kit for Staphylococcus aureus in milk and raw milk that employs a unique combination of PCR-based detection and alpha-toxin gene-specific primers. The kit utilizes a combination of alpha-toxin gene-specific primers and a universal PCR reaction mix to directly detect pathogenic Staphylococcus aureus in milk samples, providing a rapid and accurate method for monitoring milk quality and detecting S. aureus contamination.

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A rapid diagnostic kit for detecting pathogens in milk, particularly for mastitis diagnosis, that enables rapid and accurate detection of key pathogens in milk samples. The kit utilizes a novel quadruple PCR-based diagnostic system that significantly reduces the time and complexity of pathogen detection compared to traditional methods. The system enables rapid and accurate detection of major pathogens such as Streptococcus agalactiae, Staphylococcus aureus, Staphylococcus aureus, E. coli, and Bacillus cereus, providing critical information for controlling mastitis transmission and ensuring milk safety.

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