[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
Droplet digital PCR(ddPCR),as the third-generation PCR technology,demonstrates significant advantages in the etiological diagnosis of infectious diseases due to its absolute quantification,ultra-high sensitivity,and m...Droplet digital PCR(ddPCR),as the third-generation PCR technology,demonstrates significant advantages in the etiological diagnosis of infectious diseases due to its absolute quantification,ultra-high sensitivity,and multiplex detection capabilities.This article reports a case of a patient with fever of unknown origin,where ddPCR rapidly confirmed a drug-resistant bacterial infection and dynamically monitored treatment efficacy.Combining literature evidence,this paper systematically elaborates on the technical principles,clinical performance,and practical value of ddPCR in febrile patients.展开更多
Brucellosis is a global public health issue that severely affects human health,Brucella melitensis is currently the predominant species in China.Brucella spondylitis is the primary cause of the debilitating and disabl...Brucellosis is a global public health issue that severely affects human health,Brucella melitensis is currently the predominant species in China.Brucella spondylitis is the primary cause of the debilitating and disabling complications[1].The lumbar vertebra was the most commonly affected site,followed by the thoracic,cervical,thoracolumbar,and lumbosacral segments,and back pain,fever,sweating,and fatigue were the most common symptoms[2].However,the diagnosis of Brucella spondylitis is challenging owing to its wide spectrum of clinical presentations,cross-reactions with other bacteria,and low strain isolation rate.Therefore,a timely and accurate diagnosis of spinal brucellosis is crucial for implementing an effective therapeutic plan and improving treatment outcomes.Droplet digital polymerase chain reaction(ddPCR)is widely used for low-abundance nucleic acid detection and is useful for diagnosing infectious diseases[3].Therefore,this study aimed to evaluate the ddPCR approach for the diagnosis of brucellosis with spondylitis to improve its clinical diagnostic capacity.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.
文摘Droplet digital PCR(ddPCR),as the third-generation PCR technology,demonstrates significant advantages in the etiological diagnosis of infectious diseases due to its absolute quantification,ultra-high sensitivity,and multiplex detection capabilities.This article reports a case of a patient with fever of unknown origin,where ddPCR rapidly confirmed a drug-resistant bacterial infection and dynamically monitored treatment efficacy.Combining literature evidence,this paper systematically elaborates on the technical principles,clinical performance,and practical value of ddPCR in febrile patients.
基金supported by the Key Research and Development Projects of the Ningxia Hui Autonomous Region(Grant no.2022BEG03161)。
文摘Brucellosis is a global public health issue that severely affects human health,Brucella melitensis is currently the predominant species in China.Brucella spondylitis is the primary cause of the debilitating and disabling complications[1].The lumbar vertebra was the most commonly affected site,followed by the thoracic,cervical,thoracolumbar,and lumbosacral segments,and back pain,fever,sweating,and fatigue were the most common symptoms[2].However,the diagnosis of Brucella spondylitis is challenging owing to its wide spectrum of clinical presentations,cross-reactions with other bacteria,and low strain isolation rate.Therefore,a timely and accurate diagnosis of spinal brucellosis is crucial for implementing an effective therapeutic plan and improving treatment outcomes.Droplet digital polymerase chain reaction(ddPCR)is widely used for low-abundance nucleic acid detection and is useful for diagnosing infectious diseases[3].Therefore,this study aimed to evaluate the ddPCR approach for the diagnosis of brucellosis with spondylitis to improve its clinical diagnostic capacity.
