[Objectives]This study aimed to evaluate the detection sensitivity of Staphylococcus aureus in dairy products utilizing the chip digital PCR(cdPCR)technique.[Methods]Specific primers and probes were designed and synth...[Objectives]This study aimed to evaluate the detection sensitivity of Staphylococcus aureus in dairy products utilizing the chip digital PCR(cdPCR)technique.[Methods]Specific primers and probes were designed and synthesized based on the conserved sequence of the heat-resistant nuclease gene nuc of S.aureus.cdPCR was employed to detect S.aureus,and the sensitivity of this technique was systematically assessed in samples exhibiting low levels of contamination.[Results]cdPCR demonstrated precise quantification when the initial concentration of the sample enrichment solution was equal to or greater than 50 CFU/mL.The detection dynamic range extended across at least five orders of magnitude,with a minimum DNA detection limit of 0.2304 pg/μL.In artificially contaminated cheese samples,the method s lower limit of quantification for detecting S.aureus was 8×10^(2) CFU/g.Regression analysis demonstrated that the gene copy number concentration measured by cdPCR exhibited a strong linear correlation with bacterial contamination concentration across a broad range.[Conclusions]The cdPCR method developed in this study demonstrates high sensitivity and robust quantitative capabilities,offering a reliable technical approach for the precise detection of low-level S.aureus contamination in dairy products.展开更多
基金Supported by Science and Technology Program of Inner Mongolia Autonomous Region"Research and Demonstration of Novel Molecular Biological Identification Technology for Multiple Source Components in Milk and Dairy Products"(2025YFSH0029).
文摘[Objectives]This study aimed to evaluate the detection sensitivity of Staphylococcus aureus in dairy products utilizing the chip digital PCR(cdPCR)technique.[Methods]Specific primers and probes were designed and synthesized based on the conserved sequence of the heat-resistant nuclease gene nuc of S.aureus.cdPCR was employed to detect S.aureus,and the sensitivity of this technique was systematically assessed in samples exhibiting low levels of contamination.[Results]cdPCR demonstrated precise quantification when the initial concentration of the sample enrichment solution was equal to or greater than 50 CFU/mL.The detection dynamic range extended across at least five orders of magnitude,with a minimum DNA detection limit of 0.2304 pg/μL.In artificially contaminated cheese samples,the method s lower limit of quantification for detecting S.aureus was 8×10^(2) CFU/g.Regression analysis demonstrated that the gene copy number concentration measured by cdPCR exhibited a strong linear correlation with bacterial contamination concentration across a broad range.[Conclusions]The cdPCR method developed in this study demonstrates high sensitivity and robust quantitative capabilities,offering a reliable technical approach for the precise detection of low-level S.aureus contamination in dairy products.
